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Calbindin-Positive Neurons Co-express Functional Markers in a Location-Dependent Manner Within the A11 Region of the Rat Brain

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Abstract

The A11 region plays a role in numerous physiological functions, including pain and locomotor activity, and consists of a variety of neurons including GABAergic, calbindin positive (Calb+), and dopaminergic (DA) neurons. However, the neurochemical nature of Calb+ neurons and their regulatory role in the A11 region remain largely unknown. In this study, we examined the kind of functional markers co-expressed in the Calb+ neurons using sections from 8-week-old rats. To examine a marker related to classical neurotransmitters, we performed in situ hybridization for vesicular glutamate transporter 2 (vGluT2) or glutamate decarboxylase (GAD) 65 and 67, in conjunction with Calb immunohistochemistry. We found cellular co-expression of Calb with vGluT2 or GAD65/67 throughout the A11 region. Nearly all Calb+/GAD65/67+ neurons were found in the rostral-middle aspect of the A11 region. In contrast, Calb+/vGluT2+ neurons were found predominantly in the middle-caudal aspect of the A11 region. For receptors and neuropeptides, we performed immunohistochemistry for androgen receptor (AR), estrogen receptors (ERα and ERβ), and calcitonin gene-related peptide (CGRP). We found that Calb+ neurons co-expressed AR in the rostral aspect of the A11 region in both male and female rats. However, we rarely find cellular co-expression of Calb with ERα or ERβ in this region. For CGRP, we found both Calb+ neurons with or without CGRP expression. These results demonstrate that Calb+ neurons co-express many functional markers. Calb+ neurons have a distinct distribution pattern and may play a variety of regulatory roles, depending on their location within the A11 region.

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Data Availability

The data that support the findings of this study and materials used in this study are available from the corresponding author upon reasonable request.

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Acknowledgements

This work was supported by Dokkyo Medical University and by the Grants-in-Aid for Scientific Research (KAKENHI, Grant Number JP18K16495) from Japan Society for the Promotion of Science (JSPS). We would like to thank Ms. Shukuko Minami for her technical assistance and Ms. Fusae Terauchi for her assistance.

Funding

This study was funded by the Grants-in-Aid for Scientific Research (KAKENHI), Grant Number JP18K16495 from Japan Society for the Promotion of Science (JSPS) for HO.

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Author notes

  1. Tsuyoshi Yamaguchi and Hidechika Ozawa contributed equally to this work.

Authors and Affiliations

  1. Department of Histology and Neurobiology, Dokkyo Medical University, School of Medicine, 880 Kitakobayashi, Mibu-machi, Shimotsuga-gun, Tochigi, 321-0293, Japan

    Tsuyoshi Yamaguchi, Hidechika Ozawa & Shuichi Ueda

  2. Department of Anesthesia and Pain Medicine, Dokkyo Medical University, School of Medicine, 880 Kitakobayashi, Mibu-machi, Shimotsuga-gun, Tochigi, 321-0293, Japan

    Hidechika Ozawa, Shigeki Yamaguchi & Shinsuke Hamaguchi

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  1. Tsuyoshi Yamaguchi
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  3. Shigeki Yamaguchi
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  4. Shinsuke Hamaguchi
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Tsuyoshi Yamaguchi and Hidechika Ozawa. The first draft of the manuscript was written by Tsuyoshi Yamaguchi and Hidechika Ozawa, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Tsuyoshi Yamaguchi.

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Yamaguchi, T., Ozawa, H., Yamaguchi, S. et al. Calbindin-Positive Neurons Co-express Functional Markers in a Location-Dependent Manner Within the A11 Region of the Rat Brain. Neurochem Res 46, 853–865 (2021). https://doi.org/10.1007/s11064-020-03217-6

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