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Article

Immunoelectron Microscopic Characterization of Vasopressin-Producing Neurons in the Hypothalamo-Pituitary Axis of Non-Human Primates by Use of Formaldehyde-Fixed Tissues Stored at −25 °C for Several Years

1
Ushimado Marine Institute (UMI), Graduate School of Natural Science and Technology, Okayama University, Ushimado, Setouchi, Okayama 701-4303, Japan
2
Department of Biological Sciences, Faculty of Science, Kanagawa University, Hiratsuka, Kanagawa 259-1293, Japan
3
Department of Anatomy, Kawasaki Medical School, Kurashiki, Okayama 701-0192, Japan
4
Department of Physiology, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan
5
Department of Physiology, Kansai Medical University, Shinmachi, Hirakata, Osaka 573-1191, Japan
6
Department of Physiology, Anatomy & Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK
*
Author to whom correspondence should be addressed.
Academic Editors: Kazunori Kageyama and Takahiro Nemoto
Int. J. Mol. Sci. 2021, 22(17), 9180; https://doi.org/10.3390/ijms22179180
Received: 5 August 2021 / Accepted: 21 August 2021 / Published: 25 August 2021
(This article belongs to the Special Issue Molecular Mechanisms Underlying Stress Response and Resilience)
Translational research often requires the testing of experimental therapies in primates, but research in non-human primates is now stringently controlled by law around the world. Tissues fixed in formaldehyde without glutaraldehyde have been thought to be inappropriate for use in electron microscopic analysis, particularly those of the brain. Here we report the immunoelectron microscopic characterization of arginine vasopressin (AVP)-producing neurons in macaque hypothalamo-pituitary axis tissues fixed by perfusion with 4% formaldehyde and stored at −25 °C for several years (4–6 years). The size difference of dense-cored vesicles between magnocellular and parvocellular AVP neurons was detectable in their cell bodies and perivascular nerve endings located, respectively, in the posterior pituitary and median eminence. Furthermore, glutamate and the vesicular glutamate transporter 2 could be colocalized with AVP in perivascular nerve endings of both the posterior pituitary and the external layer of the median eminence, suggesting that both magnocellular and parvocellular AVP neurons are glutamatergic in primates. Both ultrastructure and immunoreactivity can therefore be sufficiently preserved in macaque brain tissues stored long-term, initially for light microscopy. Taken together, these results suggest that this methodology could be applied to the human post-mortem brain and be very useful in translational research. View Full-Text
Keywords: vasopressin; corticotrophin-releasing factor; glutamate; paraventricular nucleus of the hypothalamus; Japanese macaque monkey; post-embedding immunoelectron microscopy; dense-cored neurosecretory vesicle vasopressin; corticotrophin-releasing factor; glutamate; paraventricular nucleus of the hypothalamus; Japanese macaque monkey; post-embedding immunoelectron microscopy; dense-cored neurosecretory vesicle
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MDPI and ACS Style

Otubo, A.; Maejima, S.; Oti, T.; Satoh, K.; Ueda, Y.; Morris, J.F.; Sakamoto, T.; Sakamoto, H. Immunoelectron Microscopic Characterization of Vasopressin-Producing Neurons in the Hypothalamo-Pituitary Axis of Non-Human Primates by Use of Formaldehyde-Fixed Tissues Stored at −25 °C for Several Years. Int. J. Mol. Sci. 2021, 22, 9180. https://doi.org/10.3390/ijms22179180

AMA Style

Otubo A, Maejima S, Oti T, Satoh K, Ueda Y, Morris JF, Sakamoto T, Sakamoto H. Immunoelectron Microscopic Characterization of Vasopressin-Producing Neurons in the Hypothalamo-Pituitary Axis of Non-Human Primates by Use of Formaldehyde-Fixed Tissues Stored at −25 °C for Several Years. International Journal of Molecular Sciences. 2021; 22(17):9180. https://doi.org/10.3390/ijms22179180

Chicago/Turabian Style

Otubo, Akito, Sho Maejima, Takumi Oti, Keita Satoh, Yasumasa Ueda, John F. Morris, Tatsuya Sakamoto, and Hirotaka Sakamoto. 2021. "Immunoelectron Microscopic Characterization of Vasopressin-Producing Neurons in the Hypothalamo-Pituitary Axis of Non-Human Primates by Use of Formaldehyde-Fixed Tissues Stored at −25 °C for Several Years" International Journal of Molecular Sciences 22, no. 17: 9180. https://doi.org/10.3390/ijms22179180

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