Neurotoxicity Research

, Volume 34, Issue 3, pp 353–362 | Cite as

Bilateral Olfactory Mucosa Damage Induces the Disappearance of Olfactory Glomerulus and Reduces the Expression of Extrasynaptic α5GABAARs in the Hippocampus in Early Postnatal Sprague Dawley Rats

  • Xiaomin Zheng
  • Liang Liang
  • Changchun Hei
  • Wenjuan Yang
  • Tingyuan Zhang
  • Kai Wu
  • Yi Qin
  • Qing Chang
Original Article


Chloroform-induced olfactory mucosal degeneration has been reported in adult rats following gavage. We used fixed-point chloroform infusions on different postnatal days (PNDs) to investigate the effects of early olfactory bilateral deprivation on the main olfactory bulbs in Sprague Dawley rats. The experimental groups included rats infused with chloroform (5 μl) or saline (sham, 5 μl) on PNDs 3 and 8, and rats not receiving infusions (control) (n = 6 in all groups). Rats receiving chloroform on PND 3 showed significant hypoevolutism when compared to those in other groups (P < 0.05). There was a complete disappearance and a significant reduction in the size of olfactory glomeruli in the PND 3 and 8 groups, respectively, when compared to the respective sham groups. Rats receiving chloroform on PND 3 had significant memory impairment (P < 0.01) and increased levels of learned helplessness (P < 0.05), as measured using the Morris water maze and tail suspension tests, respectively. GABAA receptor alpha5 subunit (α5GABAAR) expression in hippocampal neurons was significantly lower in rats receiving chloroform on PND 3 than in rats in other groups (P < 0.01), as measured using immunohistochemistry and polymerase chain reaction. There was thus a critical period for the preservation of regenerative ability in olfactory receptor neurons, during which damage and olfactory deprivation led to altered rhinencephalon structure and disappearance of olfactory glomeruli, which induced hypoevolutism. Olfactory deprivation after the critical period had no significant effect on olfactory receptor neuron regeneration, leading to reduced developmental and behavioral effects in Sprague Dawley rats.


Olfactory mucosa Olfactory glomerulus α5GABAARs Sprague Dawley Development 



We thank the Esa Korpi laboratory members, for their valuable comments on this manuscript. This work is financially supported by Natural Science Foundation China (30960114, 81460644), and West China First-Class Discipline Construction Project in Basic Medicine funded by Ningxia Medical University (NXYLXK2017B07); Thank to the experimental animals’ center of Ningxia Medical University for providing the necessary artificial feeding for suckling rats.

Author Contributions

Zheng Xiaomin and Liang Liang together conceived and designed the experiments, then performed most of the experiments.

Hei Changchun, Yang Wenjuan, and Zhang Tingyuan participate in part of the experiments.

Chang Qing, Zheng Xiaomin, and Liang Liang analyzed the data.

Zheng Xiaomin, Liang Liang, Wu Kai, and QinYi contributed the reagents/materials/tools for analysis.

Zheng Xiaomin drafted the paper.

Compliance with Ethical Standards

Competing Interests

The authors declare that they have no competing interests.

Supplementary material

12640_2018_9893_MOESM1_ESM.xls (110 kb)
ESM 1 (XLS 109 kb)


  1. Andres K (1966) Der Feinbau der Regio olfactoria von Makrosmatikern. Z Zellforsch Mikrosk Anat 69:140–154CrossRefGoogle Scholar
  2. Angely CJ, Coppola DM (2010) How does long-term odor deprivation affect the olfactory capacity of adult mice? Behav Brain Funct 25(6):26CrossRefGoogle Scholar
  3. Beites CL, Kawauchi S, Crocker CE, Calof AL (2005) Identification and molecular regulation of neural stem cells in the olfactory epithelium. Exp Cell Res 306(2):309–316CrossRefGoogle Scholar
  4. Belluscio L, Katz LC (2001) Symmetry, stereotypy, and topography of odorant representations in mouse olfactory bulb. J Neurosci 21:2113–2122CrossRefGoogle Scholar
  5. Beracochea D, Celerier A, Pierard C (2004) βCCM but not physostigmine enhancement of memory retrieval depends on emotional processes in mice. J Psychopharmacol 176(1):66–73CrossRefGoogle Scholar
  6. Bodyak N, Slotnick B (1999) Performance of mice in an automated olfactometer: odor detection, discrimination and odor memory. Chem Senses 24:637–645CrossRefGoogle Scholar
  7. Brailoiu GC, Dun SL, Brailoiu E, Inan S, Yang J, Chang JK, Dun NJ (2007) Nesfatin-1: distribution and interaction with a G protein-coupled receptor in the rat brain. Endocrinology 148(10):5088–5094CrossRefGoogle Scholar
  8. Brann JH, Ellis DP, Ku BS, Spinazzi EF, Firestein S (2015) Injury in aged animals robustly activates quiescent olfactory neural stem cells. Front Neurosci 9:367–375CrossRefGoogle Scholar
  9. Brunjesa PC, Borrora MJ (1983) Unilateral odor deprivation: differential effects due to time of treatment. Brain Res Bull 11(5):501–503CrossRefGoogle Scholar
  10. Calof AL, Bonnin A, Crocker C, Kawauchi S, Murray RC, Shou J, Wu HH (2002) Progenitor cells of the olfactory receptor neuron lineage. Microsc Res Tech 58:176–188CrossRefGoogle Scholar
  11. Cavallin MA, Powell K, Biju KC, Fadool DA (2010) State-dependent sculpting of olfactory sensory neurons is attributed to sensory enrichment, odor deprivation, and aging. Neurosci Lett 483(2):90–95CrossRefGoogle Scholar
  12. Chloroform (PDF), CICAD, 58, World Health Organization (2004)Google Scholar
  13. Conover JC, Notti RQ (2008) The neural stem cell niches. Cell Tissue Res 331:211–224CrossRefGoogle Scholar
  14. Cummings DM, Brunjes PC (1997) The effects of variable periods of functional deprivation on olfactory bulb development in rats. Exp Neurol 148(1):360–366CrossRefGoogle Scholar
  15. Dorman DC, Miller KL, D'Antonio A, James RA, Morgan KT (1997) Chloroform-induced olfactory mucosal degeneration and osseous ethmoid hyperplasia are not associated with olfactory deficits in Fischer 344 rats. Toxicology 122(1–2):39–50CrossRefGoogle Scholar
  16. Ferrando S, Gallus L, Gambardella C, Ghigliotti L (2010) Cell proliferation and apoptosis in the olfactory epithelium of the shark Scyliorhinus canicula. J Chem Neuroanat 40(4):293–300CrossRefGoogle Scholar
  17. Filho CB, Jesse CR, Donato F, Del Fabbro L, de Gomes MG, Goes AT, Souza LC, Giacomeli R, Antunes M, Luchese C, Roman SS, Boeira SP (2016) Neurochemical factors associated with the antidepressant-like effect of flavonoid chrysin in chronically stressed mice. Eur J Pharmacol 791:284–296CrossRefGoogle Scholar
  18. FU Li-zhi XU, JIN-yi WU, Xiao-ming et al (2007) The advances of the γ-amino butyric acid receptors and relative drugs [J]. China Healthc Innov 02(8):29–35Google Scholar
  19. Getova DP, Dimitrova DD (2007) Effects of GABAB receptor antagonists CGP63360,CGP76290A and CGP76291A on learning and memory processes in rodents. Cent Eur J Med 2(3):280–293Google Scholar
  20. Glykys J, Mann EO, Mody I (2008) Which GABA(A) receptor subunits are necessary for tonic inhibition in the hippocampus? J Neurosci 28(6):1421–1426CrossRefGoogle Scholar
  21. Gomez CR, Plackett TP, Kovacs EJ (2007) Aging and estrogen: modulation of inflammatory responses after injury. Exp Gerontol 42(5):451–456CrossRefGoogle Scholar
  22. Graziadei PPC (1973) Cell dynamics in the olfactory mucosa. Tissue Cell 5:113–131CrossRefGoogle Scholar
  23. Graziadei P, Monti Gaziadei G (1978) Continuous nerve cell renewal in the olfactory system. Springer Verlag, New YorkCrossRefGoogle Scholar
  24. Kikuta S, Sakamoto T, Nagayama S, Kan aya K, Kinoshita M, Kondo K, Tsunoda K, Mori K, Yamasoba T (2015) Sensory deprivation disrupts homeostatic regeneration of newly generated olfactory sensory neurons after injury in adult mice. J Neurosci 35(6):2657–2673CrossRefGoogle Scholar
  25. Liang L, Xiaomin Z, Chengjun Z et al (2014) Impact of early olfactory deprivation on expression of GABAA receptor δ subunit in dentate gyrus of SD rats [J]. J Ningxia Med Univ. (16):13–18Google Scholar
  26. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408CrossRefGoogle Scholar
  27. Locci A, Porcu P, Talani G, Santoru F, Berretti R, Giunti E, Licheri V, Sanna E, Concas A (2016) Neonatal estradiol exposure to female rats changes GABAA receptor expression and function, and spatial learning during adulthood. Horm Behav 18(87):35–46Google Scholar
  28. Mackay-Sim A, Kittel P (1991) Cell dynamics in the adult mouse olfactory epithelium: a quantitative autoradiographic study. J Neurosci 11:979–984CrossRefGoogle Scholar
  29. Marcucci F (2011) Axon development and synapse formation in olfactory sensory neurons. Doctor of Philosophy thesis. Columbia UniversityGoogle Scholar
  30. Moulton DG, Celebi G, Fink RP (1970) Olfaction in mammals—two aspects: proliferation of cells in the olfactory epithelium and sensitivity to odours. In: Wolstenholme GEW, Knight J (eds) Ciba foundation on taste and smell in vertebrates. J. & A. Churchill, London, pp 227–250Google Scholar
  31. Mumm JS, Shou J, Calof AL (1996) Colony-forming progenitors from mouse olfactory epithelium: evidence for feedback regulation of neuron production. Proc Natl Acad Sci U S A 93:11167–11172CrossRefGoogle Scholar
  32. Murrell W, Bushell GR, Livesey J, McGrath J, MacDonald KP, Bates PR, Mackay-Sim A (1996) Neurogenesis in adult human. Neuroreport 7:1189–1194CrossRefGoogle Scholar
  33. Nusser Z, Mody I (2002) Selective modulation of tonic and phasic inhibitions in dentate gyrus granule cells. J Neurophysiol 87:2624–2628CrossRefGoogle Scholar
  34. Pisu MG, Dore R, Mostallino MC, Loi M, Pibiri F, Mameli R, Cadeddu R, Secci PP, Serra M (2011) Down-regulation of hippocampal BDNF and arc associated with improvement in aversive spatial memory performance in socially isolated rats. Behav Brain Res 222:73–80CrossRefGoogle Scholar
  35. Ramírez-Rodríguez G, Klempin F, Babu H, Benítez-King G, Kempermann G (2009) Melatonin modulates cell survival of new neurons in the hippocampus of adult mice. Neuropsychopharmacology 34(9):2180–2191CrossRefGoogle Scholar
  36. Santoru F, Berretti R, Locci A, Porcu P, Concas A (2014) Decreased allopregnanolone induced by hormonal contraceptives is associated with a reduction in social behavior and sexual motivation in female rats. Psychopharmacology 231:3351–3364CrossRefGoogle Scholar
  37. Schwob JE, Jang W, Holbrook EH, Lin B, Herrick DB, Peterson JN, The CJH (2017) Stem and progenitor cells of the mammalian olfactory epithelium: taking poietic license. J Comp Neurol 525(4):1034–1054CrossRefGoogle Scholar
  38. Serafini R, Maric D, Maric I, Ma W, Fritschy JM, Zhang L, Barker JL (1998) Dominant GABA(A) receptor/Cl-channel kinetics correlate with the relative expressions of alpha2, alpha3, alpha5 and beta3 subunits in embryonic rat neurones. Eur J Neurosci 10(1):334–349CrossRefGoogle Scholar
  39. Shou J, Murray RC, Rim PC, Calof AL (2000) Opposing effects of bone morphogenetic proteins on neuron production and survival in the olfactory receptor neuron lineage. Development 127:5403–5413PubMedGoogle Scholar
  40. Smart IHM (1971) Location and orientation of mitotic figures in the developing mouse olfactory epithelium. J Anat 109:243–251PubMedPubMedCentralGoogle Scholar
  41. Smith CG (1951) Regeneration of sensory epithelium and nerves in adult frogs. Anat Rec 109:661–671CrossRefGoogle Scholar
  42. Steru L, Chermat R, Thierry B, Simon P (1985) The tail suspension test: a new method for screening antidepressants in mice. Psychopharmacology 85:367–370CrossRefGoogle Scholar
  43. Suzuki Y (2007) Apoptosis and the insulin-like growth factor family in the developing olfactory epithelium. Anat Sci Int 82(4):200–206Google Scholar
  44. Whissell PD, Rosenzweig S, Lecker I et al (2013) γ-aminobutyric acid type a receptors that contain the δ subunit promote memory and neurogenesis in the dentate gyrus [J]. Ann Neurol 74(4):611–621CrossRefGoogle Scholar
  45. Wolozin B, Sunderland T, Zheng BB, Resau J, Dufy B, Barker J, Swerdlow R, Coon H (1992) Continuous culture of neuronal cells from adult human olfactory epithelium. J Mol Neurosci 3:137–146CrossRefGoogle Scholar
  46. Yang Y, Lin P, Chen F, Wang A, Lan X, Song Y, Jin Y (2013) Luman recruiting factor regulates endoplasmic reticulum stress in mouse ovarian granulosa cell apoptosis. Theriogenology 79(4):633–639CrossRefGoogle Scholar
  47. Zheng X, Shen X, Wu K et al (2011) Impact of early olfactory deprivation on the development of SD rat. Chinese J Anat 34(1):8–12Google Scholar
  48. Zou D-J, Feinstein P, Rivers AL et al (2004) Postnatal refinement of peripheral olfactory projections. Science 304(5679):1976–1979CrossRefGoogle Scholar
  49. Zunji K, Zhibin Y (2000) Change of GABAergic neurons in hippocampus of aged learning and memory impaired rats [J]. Prog Anat Sci 6(2):148–151Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Xiaomin Zheng
    • 1
  • Liang Liang
    • 2
  • Changchun Hei
    • 1
  • Wenjuan Yang
    • 3
  • Tingyuan Zhang
    • 4
  • Kai Wu
    • 1
  • Yi Qin
    • 1
  • Qing Chang
    • 1
  1. 1.Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, Key Laboratory of Reproduction and Genetic Heredity of Ningxia Hui Autonomous RegionThe School of Basic Medicine, Department of Anatomy, Ningxia Medical UniversityYinchuanChina
  2. 2.Hubei General HospitalWuhanChina
  3. 3.Tongxin County HospitalTongxin County, Ningxia Hui Autonomous RegionYinchuanChina
  4. 4.People’ s Hospital of Heze CityShandongChina

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