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The Pivotal Role of Neuropeptide Crosstalk from Ventromedial-PACAP to Dorsomedial-Galanin in the Appetite Regulation in the Mouse Hypothalamus

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Abstract

We have previously shown that pituitary adenylate cyclase-activating polypeptide (PACAP) in the ventromedial hypothalamus (VMH) enhances feeding during the dark cycle and after fasting, and inhibits feeding during the light cycle. On the other hand, galanin is highly expressed in the hypothalamus and has been reported to be involved in feeding regulation. In this study, we investigated the involvement of the VMH-PACAP to the dorsomedial hypothalamus (DMH)-galanin signaling in the regulation of feeding. Galanin expression in the hypothalamus was significantly increased with fasting, but this increment was canceled in PACAP-knockout (KO) mice. Furthermore, overexpression of PACAP in the VMH increased the expression of galanin, while knockdown (KD) of PACAP in the VMH decreased the expression of galanin, indicating that the expression of galanin in the hypothalamus might be regulated by PACAP in the VMH. Therefore, we expressed the synaptophysin-EGFP fusion protein (SypEGFP) in PACAP neurons in the VMH and visualized the neural projection to the hypothalamic region where galanin was highly expressed. A strong synaptophysin-EGFP signal was observed in the DMH, indicating that PACAP-expressing cells of the VMH projected to the DMH. Furthermore, galanin immunostaining in the DMH showed that galanin expression was weak in PACAP-KO mice. When galanin in the DMH was knocked down, food intake during the dark cycle and after fasting was decreased, and food intake during the light cycle was increased, as in PACAP-KO mice. These results indicated that galanin in the DMH may regulate the feeding downstream of PACAP in the VMH.

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Data Availability

All data generated during this study are available upon reasonable request from the corresponding authors.

Abbreviations

AAV 2/9:

AAV vector serotype 9

AgRP:

Agouti-related peptide

ANOVA:

Analysis of variance

ARC:

Arcuate nucleus

cDNA:

Complementary DNA

DMH:

Dorsomedial hypothalamus

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

KD:

Knockdown

KO:

Knockout

MCH:

Melanin-containing hormone

PACAP:

Pituitary adenylate cyclase-activating polypeptide

POMC:

Pro-opiomelanocortin

PVH:

Paraventricular hypothalamus

qPCR:

Quantitative polymerase chain reaction

RT:

Reverse transcription

SypEGFP:

Synaptophysin-EGFP fusion protein

VMH:

Ventromedial hypothalamus

WT:

Wild-type

References

  1. Muller TD, Bluher M, Tschop MH, DiMarchi RD (2021) Anti-obesity drug discovery: advances and challenges. Nat Rev Drug Discov. https://doi.org/10.1038/s41573-021-00337-8

    Article  Google Scholar 

  2. Nguyen TT, Kambe Y, Kurihara T, Nakamachi T, Shintani N, Hashimoto H, Miyata A (2020) Pituitary adenylate cyclase-activating polypeptide in the ventromedial hypothalamus Is responsible for food intake behavior by modulating the expression of agouti-related peptide in mice. Mol Neurobiol 57(4):2101–2114. https://doi.org/10.1007/s12035-019-01864-7

    Article  CAS  Google Scholar 

  3. Wang R, Yuan J, Zhang C, Wang L, Liu Y, Song L, Zhong W, Chen X et al (2018) Neuropeptide Y-positive neurons in the dorsomedial hypothalamus are involved in the anorexic effect of angptl8. Front Mol Neurosci 11:451. https://doi.org/10.3389/fnmol.2018.00451

    Article  CAS  Google Scholar 

  4. Jeong JH, Lee DK, Jo YH (2017) Cholinergic neurons in the dorsomedial hypothalamus regulate food intake. Mol Metab 6(3):306–312. https://doi.org/10.1016/j.molmet.2017.01.001

    Article  CAS  Google Scholar 

  5. Liao GY, Kinney CE, An JJ, Xu B (2019) TrkB-expressing neurons in the dorsomedial hypothalamus are necessary and sufficient to suppress homeostatic feeding. Proc Natl Acad Sci U S A 116(8):3256–3261. https://doi.org/10.1073/pnas.1815744116

    Article  CAS  Google Scholar 

  6. Imoto D, Yamamoto I, Matsunaga H, Yonekura T, Lee ML, Kato KX, Yamasaki T, Xu S et al (2021) Refeeding activates neurons in the dorsomedial hypothalamus to inhibit food intake and promote positive valence. Mol Metab 54:101366. https://doi.org/10.1016/j.molmet.2021.101366

    Article  CAS  Google Scholar 

  7. Rust VA, Crosby KM (2021) Cholecystokinin acts in the dorsomedial hypothalamus of young male rats to suppress appetite in a nitric oxide-dependent manner. Neurosci Lett 764:136295. https://doi.org/10.1016/j.neulet.2021.136295

    Article  CAS  Google Scholar 

  8. Faber CL, Deem JD, Phan BA, Doan TP, Ogimoto K, Mirzadeh Z, Schwartz MW, Morton GJ (2021) Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism. Elife 10:e63671. https://doi.org/10.7554/eLife.63671

    Article  CAS  Google Scholar 

  9. Parker JA, Bloom SR (2012) Hypothalamic neuropeptides and the regulation of appetite. Neuropharmacology 63(1):18–30. https://doi.org/10.1016/j.neuropharm.2012.02.004

    Article  CAS  Google Scholar 

  10. Fang P, Yu M, Guo L, Bo P, Zhang Z, Shi M (2012) Galanin and its receptors: a novel strategy for appetite control and obesity therapy. Peptides 36(2):331–339. https://doi.org/10.1016/j.peptides.2012.05.016

    Article  CAS  Google Scholar 

  11. Marcos P, Covenas R (2021) Neuropeptidergic control of feeding: focus on the galanin family of peptides. Int J Mol Sci 22(5):2544. https://doi.org/10.3390/ijms22052544

    Article  CAS  Google Scholar 

  12. Akabayashi A, Koenig JI, Watanabe Y, Alexander JT, Leibowitz SF (1994) Galanin-containing neurons in the paraventricular nucleus: a neurochemical marker for fat ingestion and body weight gain. Proc Natl Acad Sci U S A 91(22):10375–10379. https://doi.org/10.1073/pnas.91.22.10375

    Article  CAS  Google Scholar 

  13. Barson JR, Morganstern I, Leibowitz SF (2010) Galanin and consummatory behavior: special relationship with dietary fat, alcohol and circulating lipids. Exp Suppl 102:87–111. https://doi.org/10.1007/978-3-0346-0228-0_8

    Article  CAS  Google Scholar 

  14. Cheung CC, Hohmann JG, Clifton DK, Steiner RA (2001) Distribution of galanin messenger RNA-expressing cells in murine brain and their regulation by leptin in regions of the hypothalamus. Neuroscience 103(2):423–432. https://doi.org/10.1016/s0306-4522(01)00012-4

    Article  CAS  Google Scholar 

  15. Hashimoto H, Shintani N, Tanaka K, Mori W, Hirose M, Matsuda T, Sakaue M, Miyazaki J et al (2001) Altered psychomotor behaviors in mice lacking pituitary adenylate cyclase-activating polypeptide (PACAP). Proc Natl Acad Sci U S A 98(23):13355–13360. https://doi.org/10.1073/pnas.231094498

    Article  CAS  Google Scholar 

  16. Harris JA, Hirokawa KE, Sorensen SA, Gu H, Mills M, Ng LL, Bohn P, Mortrud M et al (2014) Anatomical characterization of Cre driver mice for neural circuit mapping and manipulation. Front Neural Circuits 8:76. https://doi.org/10.3389/fncir.2014.00076

    Article  Google Scholar 

  17. Kambe Y, Thi TN, Hashiguchi K, Sameshima Y, Yamashita A, Kurihara T, Miyata A (2022) The dorsal hippocampal protein targeting to glycogen maintains ionotropic glutamate receptor subunits expression and contributes to working and short-term memories in mice. J Pharmacol Sci 148(1):108–115. https://doi.org/10.1016/j.jphs.2021.10.008

    Article  CAS  Google Scholar 

  18. Sarbassov DD, Guertin DA, Ali SM, Sabatini DM (2005) Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science 307(5712):1098–1101. https://doi.org/10.1126/science.1106148

    Article  CAS  Google Scholar 

  19. Oh SW, Harris JA, Ng L, Winslow B, Cain N, Mihalas S, Wang Q, Lau C et al (2014) A mesoscale connectome of the mouse brain. Nature 508(7495):207–214. https://doi.org/10.1038/nature13186

    Article  CAS  Google Scholar 

  20. Aurnhammer C, Haase M, Muether N, Hausl M, Rauschhuber C, Huber I, Nitschko H, Busch U et al (2012) Universal real-time PCR for the detection and quantification of adeno-associated virus serotype 2-derived inverted terminal repeat sequences. Hum Gene Ther Methods 23(1):18–28. https://doi.org/10.1089/hgtb.2011.034

    Article  CAS  Google Scholar 

  21. Kirihara Y, Takechi M, Kurosaki K, Kobayashi Y, Kurosawa T (2013) Anesthetic effects of a mixture of medetomidine, midazolam and butorphanol in two strains of mice. Exp Anim 62(3):173–180. https://doi.org/10.1538/expanim.62.173

    Article  CAS  Google Scholar 

  22. Paxinos G, Franklin KBJ (2001) The mouse brain in stereotaxic coordinates. Academic Press

    Google Scholar 

  23. Rodgers RJ, Ishii Y, Halford JC, Blundell JE (2002) Orexins and appetite regulation. Neuropeptides 36(5):303–325. https://doi.org/10.1016/s0143-4179(02)00085-9

    Article  CAS  Google Scholar 

  24. Hohmann JG, Krasnow SM, Teklemichael DN, Clifton DK, Wynick D, Steiner RA (2003) Neuroendocrine profiles in galanin-overexpressing and knockout mice. Neuroendocrinology 77(6):354–366. https://doi.org/10.1159/000071308

    Article  CAS  Google Scholar 

  25. Adams AC, Clapham JC, Wynick D, Speakman JR (2008) Feeding behaviour in galanin knockout mice supports a role of galanin in fat intake and preference. J Neuroendocrinol 20(2):199–206. https://doi.org/10.1111/j.1365-2826.2007.01638.x

    Article  CAS  Google Scholar 

  26. Tachibana T, Mori M, Khan MS, Ueda H, Sugahara K, Hiramatsu K (2008) Central administration of galanin stimulates feeding behavior in chicks. Comp Biochem Physiol A Mol Integr Physiol 151(4):637–640. https://doi.org/10.1016/j.cbpa.2008.08.001

    Article  CAS  Google Scholar 

  27. Chen KS, Xu M, Zhang Z, Chang WC, Gaj T, Schaffer DV, Dan Y (2018) A hypothalamic switch for REM and non-REM sleep. Neuron 97 (5):1168–1176 e1164. https://doi.org/10.1016/j.neuron.2018.02.005

  28. Patterson M, Murphy KG, Thompson EL, Smith KL, Meeran K, Ghatei MA, Bloom SR (2006) Microinjection of galanin-like peptide into the medial preoptic area stimulates food intake in adult male rats. J Neuroendocrinol 18(10):742–747. https://doi.org/10.1111/j.1365-2826.2006.01473.x

    Article  CAS  Google Scholar 

  29. Lawrence CB, Williams T, Luckman SM (2003) Intracerebroventricular galanin-like peptide induces different brain activation compared with galanin. Endocrinology 144(9):3977–3984. https://doi.org/10.1210/en.2003-0391

    Article  CAS  Google Scholar 

  30. Engel JA, Hjorth S, Svensson K, Carlsson A, Liljequist S (1984) Anticonflict effect of the putative serotonin receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT). Eur J Pharmacol 105(3–4):365–368. https://doi.org/10.1016/0014-2999(84)90634-4

    Article  CAS  Google Scholar 

  31. Bing O, Moller C, Engel JA, Soderpalm B, Heilig M (1993) Anxiolytic-like action of centrally administered galanin. Neurosci Lett 164(1–2):17–20. https://doi.org/10.1016/0304-3940(93)90846-d

    Article  CAS  Google Scholar 

  32. Holmes A, Kinney JW, Wrenn CC, Li Q, Yang RJ, Ma L, Vishwanath J, Saavedra MC et al (2003) Galanin GAL-R1 receptor null mutant mice display increased anxiety-like behavior specific to the elevated plus-maze. Neuropsychopharmacology 28(6):1031–1044. https://doi.org/10.1038/sj.npp.1300164

    Article  CAS  Google Scholar 

  33. Karlsson RM, Holmes A (2006) Galanin as a modulator of anxiety and depression and a therapeutic target for affective disease. Amino Acids 31(3):231–239. https://doi.org/10.1007/s00726-006-0336-8

    Article  CAS  Google Scholar 

  34. Lehmann ML, Mustafa T, Eiden AM, Herkenham M, Eiden LE (2013) PACAP-deficient mice show attenuated corticosterone secretion and fail to develop depressive behavior during chronic social defeat stress. Psychoneuroendocrinology 38(5):702–715. https://doi.org/10.1016/j.psyneuen.2012.09.006

    Article  CAS  Google Scholar 

  35. Vaudry D, Falluel-Morel A, Bourgault S, Basille M, Burel D, Wurtz O, Fournier A, Chow BK et al (2009) Pituitary adenylate cyclase-activating polypeptide and its receptors: 20 years after the discovery. Pharmacol Rev 61(3):283–357. https://doi.org/10.1124/pr.109.001370

    Article  CAS  Google Scholar 

  36. Joo KM, Chung YH, Kim MK, Nam RH, Lee BL, Lee KH, Cha CI (2004) Distribution of vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide receptors (VPAC1, VPAC2, and PAC1 receptor) in the rat brain. J Comp Neurol 476(4):388–413. https://doi.org/10.1002/cne.20231

    Article  CAS  Google Scholar 

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Acknowledgements

We would like to thank Ms. Izumi Fujishima for their technical contribution and all the Joint Research Laboratory and the Division of Laboratory Animal Sciences, Kagoshima University, to help with animal care and the use of the facilities. The data in the present manuscript have been presented at the 95th annual meeting of the Japanese Society for Pharmacology (https://www.jstage.jst.go.jp/article/jpssuppl/95/0/95_1-P-004/_article/-char/ja/.). Still, no figures contained in the present paper have been published in a journal before.

Funding

We would like to express our deepest gratitude for a grant from the Kodama Memorial Fund for Medical Research for their research support and a grant from the Takeda Science Foundation. This work was supported by JSPS KAKENHI grant numbers JP20K07067, JP20H03429, JP18K07394, JP21K19335, JP20H00492, and JP19K07121; MEXT KAKENHI grant number JP18H05416; AMED grant number JP21dm0207117.

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Authors and Affiliations

  1. Department of Pharmacology, Graduate School of Medical and Dental Science, Kagoshima University, Sakuragaoka 8-35-1, Kagoshima-shi, Kagoshima, 890-8544, Japan

    Yuki Kambe, Thanh Trung Nguyen, Thu Thi Nguyen, Yoshimune Sameshima, Kohei Hashiguchi, Takashi Kurihara & Atsuro Miyata

  2. Department of Health and Nutrition, Faculty of Health Sciences, University of Health and Welfare, Shimamicho 1398, Kita-ku, Niigata, 950-3198, Japan

    Toshiharu Yasaka

  3. Laboratory of Pharmacology, School of Pharmaceutical Sciences, Wakayama Medical University, 25-1 Shichibancho, Wakayama-shi, Wakayama, 640-8156, Japan

    Norihito Shintani & Hitoshi Hashimoto

  4. Laboratory of Molecular Neuropharmacology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Norihito Shintani

  5. United Graduate School of Child Development, Osaka University, Kanazawa University and Hamamatsu University School of Medicine, Chiba University and University of Fukui, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Hitoshi Hashimoto

  6. Division of Bioscience, Institute for Datability Science, Osaka University, 2-8 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Hitoshi Hashimoto

  7. Transdimensional Life Imaging Division, Institute for Open and Transdisciplinary Research Initiatives, Osaka University, 1-1 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Hitoshi Hashimoto

  8. Department of Molecular Pharmaceutical Sciences, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Hitoshi Hashimoto

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  1. Yuki Kambe
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Contributions

YK carried out the experiments, performed behavioral studies, performed the statistical analysis, and wrote the manuscript. TTN, YS, and KH carried out the experiments. TY, TK, and AM conceived of and participated in the study’s design and wrote the manuscript. NS and HH participated in the study’s design and reviewed the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Yuki Kambe.

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Ethics Approval

All experiments in the present study were approved by the Experimental Animal Research Committee of Kagoshima University (Approval numbers: MD16076, MD16116, and MD17054) and the Gene Recombination Experiment Safety Committee of Kagoshima University (Approval numbers: S28006, S28015, and S29007). All animal experiments were performed following the ARRIVE guidelines and the National Institutes of Health guide for the care and use of laboratory animals. The work described in the present study was carried out following The Code of Ethics of the World Medical Association for animal experiments (http://ec.europa.eu/environment/chemicals/lab_animals/legislation_en.htm) and Uniform Requirements for manuscripts submitted to Biomedical Journals (http://www.icmje.org).

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Kambe, Y., Nguyen, T.T., Yasaka, T. et al. The Pivotal Role of Neuropeptide Crosstalk from Ventromedial-PACAP to Dorsomedial-Galanin in the Appetite Regulation in the Mouse Hypothalamus. Mol Neurobiol 60, 171–182 (2023). https://doi.org/10.1007/s12035-022-03084-y

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