Age-Related Changes in Ang II Receptor Localization and Expression in the Developing Auditory Pathway

  • M. E. Arce
  • S. I. Sánchez
  • M. M. Correa
  • G. M. CiuffoEmail author
Original Paper


We studied Ang II receptor localization in different nuclei of the auditory system, by means of binding autoradiography, during brain development. The inferior colliculus (IC), a large midbrain structure which serves as an obligatory synaptic station in both the ascending and descending auditory pathways, exhibited high Ang II AT2 binding at all ages (P0, P8, P15, P30), being maximal at P15. These observations were confirmed by in situ hybridization and immunofluorescence at P15, demonstrating that AT2 receptor mRNA localized at the same area recognized by AT2 antibodies and anti β III–tubulin suggesting the neuronal nature of the reactive cells. Ang II AT1 receptors were absent at early developmental ages (P0) in all nuclei of the auditory system and a low level was observed in the IC at the age P8. AT2 receptors were present at ventral cochlear nucleus and superior olivary complex, being higher at P15 and P8, respectively. We also explored the effect of prenatal administration of Ang II or PD123319 (AT2 antagonist) on binding of Ang II receptors at P0, P8, P15. Both treatments increased significantly the level of AT2 receptors at P0 and P8 in the IC. Although total binding in the whole IC from P15 animals showed no difference between treatments, the central nucleus of the IC exhibited higher binding. Our results supports a correlation between the timing of the higher expression of Ang II AT2 receptors in different nuclei, the onset of audition and the establishment of neuronal circuits of the auditory pathway.


AT2 receptors Inferior colliculus Superior olivary complex Cochlear nucleus Brain development 



GM Ciuffo is a member of the Researcher career from CONICET. This work was supported by grants from CONICET, PIP 656 (GM Ciuffo), PICT 15-32350 (ANPCYT) (GMC), Universidad Nacional de San Luis, PROICO 02-0616 (GMC), Argentina.

Supplementary material

11064_2018_2687_MOESM1_ESM.tif (4.6 mb)
Immunolabeling of AT2 receptors at the IC of P15 rat brains. a, b. Peroxidase-DAB immunoreactivity of AT2 receptor in coronal section of P15 rat IC at low (a) and higher (b) magnification. Arrows indicate AT2 immunoreactive cell. Scale bar: 100 µm. c. Immunofluorescent staining of AT2 receptor in coronal section of P15 rat IC. Arrows indicate AT2 immunoreactive cell. Scale bar: 100 µm. d. Immunofluorescent staining of βIII-tubulin in coronal section of P15 rat IC (parallel to c). Scale bar: 100 µm. (TIF 4734 KB)
11064_2018_2687_MOESM2_ESM.tif (2.7 mb)
Detection of AT2 receptor mRNA in the IC of P15 rat brains by in situ hybridization. a. H&E stained IC section from P15 rat brain. CIC: central nucleus of the IC; 2: area 2 of Cerebellum. b. Antisense digoxigenin AT2 riboprobe labeling of the IC. c. Sense digoxigenin AT2 riboprobe labeling of the IC (negative control). Scale bar: 100 µm. (TIF 2770 KB)
11064_2018_2687_MOESM3_ESM.tif (634 kb)
Quantification of binding autoradiography (Fig. <link rid="fig3">3</link>) in control and treated animals at the stages P0, P8 and P15. Binding densities were analyzed as indicated under Methods. Total binding, binding to AT1 receptors and AT2 receptors were obtained for the different treatments and developmental stages. Data are mean ± SEM from 4 independent treatments of pregnant mothers (20–24 animals from each treatment). ***p<0.001, **p<0.01, *p<0.05 vs. control. (TIF 635 KB)


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Copyright information

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

Authors and Affiliations

  1. 1.Instituto Multidisciplinario de Investigaciones Biológicas (IMIBIO-SL CONICET)Universidad Nacional de San LuisSan LuisArgentina

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