Expression of the c-fos-monomeric red fluorescent protein 1 fusion gene in the spinal cord and the hypothalamic paraventricular nucleus in transgenic rats after nociceptive stimulation

2012 ◽  
Vol 1479 ◽  
pp. 52-61 ◽  
Author(s):  
Toru Ishikura ◽  
Hitoshi Suzuki ◽  
Mitsuhiro Yoshimura ◽  
Jun-ichi Ohkubo ◽  
Akiko Katoh ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Paraventricular Nucleus ◽  
Fluorescent Protein ◽  
Fusion Gene ◽  
Hypothalamic Paraventricular Nucleus ◽  
Red Fluorescent Protein ◽  
Nociceptive Stimulation ◽  
Transgenic Rats

scholarly journals Highly Visible Expression of an Oxytocin-Monomeric Red Fluorescent Protein 1 Fusion Gene in the Hypothalamus and Posterior Pituitary of Transgenic Rats

Endocrinology ◽  
2011 ◽  
Vol 152 (7) ◽  
pp. 2768-2774 ◽  
Author(s):  
Akiko Katoh ◽  
Hiroaki Fujihara ◽  
Toyoaki Ohbuchi ◽  
Tatsushi Onaka ◽  
Takashi Hashimoto ◽  
...  
Keyword(s):  
Median Eminence ◽  
Fluorescent Protein ◽  
Fusion Gene ◽  
Female Rats ◽  
Posterior Pituitary ◽  
Red Fluorescent Protein ◽  
Protein Fusion ◽  
Transgenic Rats ◽  
Salt Loading ◽  
Male And Female

We have generated rats bearing an oxytocin (OXT)-monomeric red fluorescent protein 1 (mRFP1) fusion transgene. The mRFP1 fluorescence was highly visible in ventral part of the supraoptic nucleus (SON) and the posterior pituitary in a whole mount. mRFP1 fluorescence in hypothalamic sections was also observed in the SON, the paraventricular nucleus (PVN), and the internal layer of the median eminence. Salt loading for 5 d caused a marked increase in mRFP1 fluorescence in the SON, the PVN, the median eminence, and the posterior pituitary. In situ hybridization histochemistry revealed that the expression of the mRNA encoding the OXT-mRFP1 fusion gene was observed in the SON and the PVN of euhydrated rats and increased dramatically after chronic salt loading. The expression of the endogenous OXT and the arginine vasopressin (AVP) genes were significantly increased in the SON and the PVN after chronic salt loading in both nontransgenic and transgenic rats. These responses were not different between male and female rats. Compared with nontransgenic rats, euhydrated and salt-loaded male and female transgenic rats showed no significant differences in plasma osmolality, sodium concentration, OXT, and AVP levels. Finally, we succeeded in generating a double-transgenic rat that expresses both the OXT-mRFP1 fusion gene and the AVP-enhanced green fluorescent protein fusion gene. Our new transgenic rats are valuable new tools to study the physiology of the hypothalamo-neurohypophysial system.

scholarly journals Specific expression of an oxytocin-enhanced cyan fluorescent protein fusion transgene in the rat hypothalamus and posterior pituitary

2009 ◽  
Vol 204 (3) ◽  
pp. 275-285 ◽  
Author(s):  
Akiko Katoh ◽  
Hiroaki Fujihara ◽  
Toyoaki Ohbuchi ◽  
Tatsushi Onaka ◽  
W Scott Young ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Fusion Gene ◽  
Plasma Osmolality ◽  
Cyan Fluorescent Protein ◽  
Posterior Pituitary ◽  
Protein Fusion ◽  
Transgenic Rats ◽  
Salt Loading ◽  
Wide Range ◽  
Enhanced Cyan Fluorescent Protein

We have generated rats bearing an oxytocin (OXT)-enhanced cyan fluorescent protein (eCFP) fusion transgene designed from a murine construct previously shown to be faithfully expressed in transgenic mice. In situ hybridisation histochemistry revealed that the Oxt–eCfp fusion gene was expressed in the supraoptic nucleus (SON) and the paraventricular nucleus (PVN) in these rats. The fluorescence emanating from eCFP was observed only in the SON, the PVN, the internal layer of the median eminence and the posterior pituitary (PP). In in vitro preparations, freshly dissociated cells from the SON and axon terminals showed clear eCFP fluorescence. Immunohistochemistry for OXT and arginine vasopressin (AVP) revealed that the eCFP fluorescence co-localises with OXT immunofluorescence, but not with AVP immunofluorescence in the SON and the PVN. Although the expression levels of the Oxt–eCfp fusion gene in the SON and the PVN showed a wide range of variations in transgenic rats, eCFP fluorescence was markedly increased in the SON and the PVN, but decreased in the PP after chronic salt loading. The expression of the Oxt gene was significantly increased in the SON and the PVN after chronic salt loading in both non-transgenic and transgenic rats. Compared with wild-type animals, euhydrated and salt-loaded male and female transgenic rats showed no significant differences in plasma osmolality, sodium concentration and OXT and AVP levels, suggesting that the fusion gene expression did not disturb any physiological processes. These results suggest that our new transgenic rats are a valuable new tool to identify OXT-producing neurones and their terminals.

Peptidergic nature of nociception-related projections from the hypothalamic paraventricular nucleus to the dorsal horn of the spinal cord

2018 ◽  
Vol 685 ◽  
pp. 124-130 ◽  
Author(s):  
Mohammed Gamal-Eltrabily ◽  
Cecilia Márquez-Morales ◽  
Guadalupe Martínez-Lorenzana ◽  
Abimael González-Hernández ◽  
Miguel Condés-Lara
Keyword(s):  
Spinal Cord ◽  
Dorsal Horn ◽  
Paraventricular Nucleus ◽  
Hypothalamic Paraventricular Nucleus

scholarly journals Presynaptic glutamatergic transmission and feedback system of oxytocinergic neurons in the hypothalamus of a rat model of adjuvant arthritis

2020 ◽  
Vol 16 ◽  
pp. 174480692094333
Author(s):  
Teruaki Fujitani ◽  
Takanori Matsuura ◽  
Makoto Kawasaki ◽  
Hitoshi Suzuki ◽  
Haruki Nishimura ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Paraventricular Nucleus ◽  
Rat Model ◽  
Adjuvant Arthritis ◽  
Fluorescent Protein ◽  
Feedback System ◽  
Red Fluorescent Protein ◽  
Excitatory Postsynaptic Currents ◽  
Postsynaptic Currents ◽  
Bath Application

The neurohypophysial hormone oxytocin (OXT) is synthesized in the hypothalamic paraventricular and supraoptic nuclei. Recently, some studies have considered OXT to be important in sensory modulation and that the OXT protein is upregulated by acute and chronic nociception. However, the mechanism by which OXT is upregulated in neurons is unknown. In this study, we examined the resting membrane potentials and excitatory postsynaptic currents in OXT-ergic neurons in the paraventricular nucleus in adjuvant arthritis rat model, a model of chronic inflammation, using whole-cell patch-clamping. Transgenic rats expressing OXT and monomeric red fluorescent protein 1 (mRFP1) fusion protein to visualize the OXT-ergic neurons were used, and the OXT-mRFP1 transgenic rat model of adjuvant arthritis was developed by injection of heat-killed Mycobacterium butyricum. Furthermore, the feedback system of synthesized OXT was also examined using the OXT receptor antagonist L-368,899. We found that the resting membrane potentials and frequency of miniature excitatory postsynaptic currents and spontaneous excitatory postsynaptic currents in OXT-monomeric red fluorescent protein 1 neurons in the paraventricular nucleus were significantly increased in adjuvant arthritis rats. Furthermore, L-368,899 dose-dependently increased the frequency of miniature excitatory postsynaptic currents and spontaneous excitatory postsynaptic currents in OXT-ergic neurons. Following bath application of the GABAA receptor antagonist picrotoxin and the cannabinoid receptor 1 antagonist AM 251, L-368,899 still increased the frequency of miniature excitatory postsynaptic currents. However, following bath application of the nitric oxide synthase inhibitor Nω-Nitro-L-arginine methyl ester hydrochloride, L-368,899 did not alter the miniature excitatory postsynaptic current frequency. Thus, it is suggested that OXT-ergic neuron activity is upregulated via an increase in glutamate release, and that the upregulated OXT neurons have a feedback system with released endogenous OXT. It is possible that nitric oxide, but not GABA, may contribute to the feedback system of OXT neurons in chronic inflammation.

Nociceptive stimulation induces the c-fos-mRFP fusion gene expression in the spinal cord and the hypothalamus of a transgenic rat

2010 ◽  
Vol 68 ◽  
pp. e164
Author(s):  
Toru Ishikura ◽  
Hitoshi Suzuki ◽  
Hiroaki Fujihara ◽  
Hideo Ohnishi ◽  
Yoichi Ueta
Keyword(s):  
Gene Expression ◽  
Spinal Cord ◽  
Fusion Gene ◽  
Transgenic Rat ◽  
Nociceptive Stimulation
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