Neuroanatomical characterization of a growth hormone secretagogue receptor-green fluorescent protein reporter mouse

Bharath K. Mani, Angela K. Walker, Eduardo J. Lopez Soto, Jesica Raingo, Charlotte E. Lee, Mario Perelló, Zane B. Andrews, Jeffrey M. Zigman

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Growth hormone secretagogue receptor (GHSR) 1a is the only molecularly identified receptor for ghrelin, mediating ghrelin-related effects on eating, body weight, and blood glucose control, among others. The expression pattern of GHSR within the brain has been assessed previously by several neuroanatomical techniques. However, inherent limitations to these techniques and the lack of reliable anti-GHSR antibodies and reporter rodent models that identify GHSR-containing neurons have prevented a more comprehensive functional characterization of ghrelin-responsive neurons. Here we have systematically characterized the brain expression of an enhanced green fluorescence protein (eGFP) transgene controlled by the Ghsr promoter in a recently reported GHSR reporter mouse. Expression of eGFP in coronal brain sections was compared with GHSR mRNA expression detected in the same sections by in situ hybridization histochemistry. eGFP immunoreactivity was detected in several areas, including the prefrontal cortex, insular cortex, olfactory bulb, amygdala, and hippocampus, which showed no or low GHSR mRNA expression. In contrast, eGFP expression was low in several midbrain regions and in several hypothalamic nuclei, particularly the arcuate nucleus, where robust GHSR mRNA expression has been well-characterized. eGFP expression in several brainstem nuclei showed high to moderate degrees of colocalization with GHSR mRNA labeling. Further quantitative PCR and electrophysiological analyses of eGFP-labeled hippocampal cells confirmed faithful expression of eGFP within GHSR-containing, ghrelin-responsive neurons. In summary, the GHSR-eGFP reporter mouse model may be a useful tool for studying GHSR function, particularly within the brainstem and hippocampus; however, it underrepresents GHSR expression in nuclei within the hypothalamus and midbrain. J. Comp. Neurol. 522:3644-3666, 2014.

Original languageEnglish (US)
Pages (from-to)3644-3666
Number of pages23
JournalJournal of Comparative Neurology
Volume522
Issue number16
DOIs
StatePublished - Nov 1 2014

Fingerprint

Ghrelin Receptor
Green Fluorescent Proteins
Fluorescence
Ghrelin
Proteins
Messenger RNA
Mesencephalon
Neurons
Brain Stem
Hippocampus
Brain
Arcuate Nucleus of Hypothalamus
Olfactory Bulb
Amygdala
Prefrontal Cortex

Keywords

  • Ghrelin
  • GHSR
  • Reporter mouse model

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Neuroanatomical characterization of a growth hormone secretagogue receptor-green fluorescent protein reporter mouse. / Mani, Bharath K.; Walker, Angela K.; Lopez Soto, Eduardo J.; Raingo, Jesica; Lee, Charlotte E.; Perelló, Mario; Andrews, Zane B.; Zigman, Jeffrey M.

In: Journal of Comparative Neurology, Vol. 522, No. 16, 01.11.2014, p. 3644-3666.

Research output: Contribution to journalArticle

Mani, Bharath K. ; Walker, Angela K. ; Lopez Soto, Eduardo J. ; Raingo, Jesica ; Lee, Charlotte E. ; Perelló, Mario ; Andrews, Zane B. ; Zigman, Jeffrey M. / Neuroanatomical characterization of a growth hormone secretagogue receptor-green fluorescent protein reporter mouse. In: Journal of Comparative Neurology. 2014 ; Vol. 522, No. 16. pp. 3644-3666.
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AB - Growth hormone secretagogue receptor (GHSR) 1a is the only molecularly identified receptor for ghrelin, mediating ghrelin-related effects on eating, body weight, and blood glucose control, among others. The expression pattern of GHSR within the brain has been assessed previously by several neuroanatomical techniques. However, inherent limitations to these techniques and the lack of reliable anti-GHSR antibodies and reporter rodent models that identify GHSR-containing neurons have prevented a more comprehensive functional characterization of ghrelin-responsive neurons. Here we have systematically characterized the brain expression of an enhanced green fluorescence protein (eGFP) transgene controlled by the Ghsr promoter in a recently reported GHSR reporter mouse. Expression of eGFP in coronal brain sections was compared with GHSR mRNA expression detected in the same sections by in situ hybridization histochemistry. eGFP immunoreactivity was detected in several areas, including the prefrontal cortex, insular cortex, olfactory bulb, amygdala, and hippocampus, which showed no or low GHSR mRNA expression. In contrast, eGFP expression was low in several midbrain regions and in several hypothalamic nuclei, particularly the arcuate nucleus, where robust GHSR mRNA expression has been well-characterized. eGFP expression in several brainstem nuclei showed high to moderate degrees of colocalization with GHSR mRNA labeling. Further quantitative PCR and electrophysiological analyses of eGFP-labeled hippocampal cells confirmed faithful expression of eGFP within GHSR-containing, ghrelin-responsive neurons. In summary, the GHSR-eGFP reporter mouse model may be a useful tool for studying GHSR function, particularly within the brainstem and hippocampus; however, it underrepresents GHSR expression in nuclei within the hypothalamus and midbrain. J. Comp. Neurol. 522:3644-3666, 2014.

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