ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2019, Vol. 50 ›› Issue (1): 159-168.doi: 10.11843/j.issn.0366-6964.2019.01.019

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The Effect of 17β-estradiol on the Ca2+ Concentration in DRG Neurons of Newborn Rabbits

GUO Yaru, ZHANG Jin, ZHANG Yingli, ZHAO Jianshuai, DU Yi'nan, XIN Xianmeng, XU Yongping*   

  1. College of Veterinary Medicine, Northwest A & F University, Yangling 712100, China
  • Received:2018-05-02 Online:2019-01-23 Published:2019-01-24

Abstract:

The study was conducted to investigate whether estrogen can affect the regulation of primary sensory neurons, and to explore the role of Ca2+ in this mechanism. The distribution of estrogen receptor (ER) in primary culture dorsal root ganglion (DRG) from newborn rabbits was observed by Immunocytofluorescent (ICF), and the effects of 17β-E2 (1, 10, 100, 1 000 nmol·L-1) on cytosolic free Ca2+ fluorescence in DRG neurons cultured for 72 h were monitored by Laser Scanning Confocal Microscopy (LSCM). It was found that, in primary cultured DRG neurons, ERα, ERβ and GPR30 were 45.61%, 32.39%, 39.82% positive or strongly positive neurons respectively, and the neurons proportions of different sizes in different ER positive or strongly positive neurons were different. ERα and ERβ were strongly positive in DRG neuron nuclei, medium positive in cytoplasm, and weakly positive in a little neurite. GPR30 were weakly positive or negative in DRG neuron nuclei, medium positive or strongly positive in cytoplasm, and weakly positive in neurite. A certain degree of "heterogeneity" of three types of ERs (ERα, ERβ, and GPR30) was observed in the number, size type, degree of response and distribution in positive neurons. According to the variation of Ca2+ fluorescence intensity induced by 17β-E2, DRG neurons were divided into 3 types, excitatory (19.28±0.70)%, inhibitory (3.54±0.02)% and insensitive neurons (77.17±1.48)%, respectively. Excitatory neurons were divided into strongly excitatory neurons (0.195±0.118) and weakly excitatory neurons (0.032±0.003) depending on the intensity of Ca2+ fluorescence changes. Interestingly, with the increase of 17β-E2, the proportion of strongly excitatory neurons decreased, but no difference was detected in the proportion of total excitatory neurons at each group. It was indicated that with the increasing of 17β-E2, the increasement of[Ca2+]i was inhibited in strongly excitatory neurons, but not completely inhibited. These results suggest that estrogen may have distinct effects on various sensory neurons; Ca2+ pathway is one of the mechanisms in the effects of estrogen on the activity of some primary sensory neurons, and estrogen inhibits the effect of Ca2+ in DRG neurons in two ways:a direct inhibition of the effect of Ca2+ signaling pathway in several neurons, and a certain inhibitory impact on the effect of Ca2+ increment in strongly excitatory neurons with the increasing estrogen.

Key words: Ca2+, 17β-estradiol, dorsal root ganglion, newborn rabbits

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