Table 2.
Selected proteomic studies in whole hypothalamus and by hypothalamic sub-region
| Study | Animal | Extraction | Target(s) a priori? | Screen [S], Validation [V] | Map or Schematic | Major findings |
|---|---|---|---|---|---|---|
| Whole hypothalamus | ||||||
| Sung et al. 2004 | Rt | dissection | N | MALDI-TOF MS | N | 36 proteins expressed in a neuropathic pain model relative to controls |
| Kuo et al. 2005 | Ck | dissection | N | 2D-GE [S] LC-MS/MS, qRT-PCR [V] | N | 6 proteins associated with high egg production |
| Roth et al. 2006 | Rt | dissection | N | 2D-LC-MS/MS & clCAT | N | Found differential expression of five proteins involved in glutamate metabolism in juvenile versus peri-pubertal females |
| Skynner et al. 2006 | Ms | dissection | N | MALDI-TOF MS | N | Chronic corticosterone altered markers of glycolysis, gluconeogenesis and nitrogen metabolism |
| Kuhla et al. 2007 | Cw | dissection | N | MALDI-TOF MS [S] | N | Found nine proteins differentially expressed in ad libitum fed vs. energy restricted cows |
| Ropp et al. 2008 | Ms | dissection | N | SELDI-TOF | N | Distinct protein profiles following sub-acute pyridostigmine treatment |
| Sarkar et al. 2008 | Ms | dissection | N | 2D-GE, WB, MALDI-TOF-MS | N | Found seven proteins that were significantly different in hypothalamus in control versus microgravity-treated animals |
| J. Y. Lee et al. 2009 | Rt | dissection | N | 2D-GE; MALDI-TOF/MS | N | Found several proteins up-regulated following lithium treatment |
| Mishra et al. 2009 | Rt | dissection | N | 2D-GE, LC-MS/MS [S]; WB [V] | N | Light-dark shifts in circadian cycle resulted in increased food intake, body weight gain, retroperitoneal fat mass, and expression levels of 4 of 5 hypothalamic 2D-GE spots; these were identified by MS and included glycolytic and citric acid cycle enzymes. |
| H.-J. Kim et al. 2010 | Rt | dissection | N | 2D-GE, MALDI-TOF MS | N | Maternal separation was associated with down-regulation of 14 proteins from hypothalamus; maternal separation with acupuncture was associated with five down- and nine up-regulated proteins relative to maternal separation alone. |
| Popesku et al. 2010 | Fs | dissection | N | iTRAQ labeling & MS | N | 42 proteins differentially regulated by treatment with DA receptor agonists |
| Argüelles et al. 2011 | Rt | NS | N | MALDI-TOF/TOF-MS | N | Oxidative stress could be involved in the alterations of eEF-2 and several other proteins |
| Q. Wang et al. 2011 | Rt | dissection | N | MALDI-TOF/TOF-MS | N | Ubiquitin was significantly decreased in diet-resistant rats but not changed in diet-induced obese rats |
| Alexandre-Gouabau et al. 2012 | Rt | dissection | N | LC-MS/MS | N | Protein restriction in utero alters numerous pathways |
| Gasperini et al. 2012 | Rt | dissection | N | 2D-GE, MALDI-TOF MS/MS | N | Found 26 of 28 protein spots on 2D gels for hypothalamus show significant expression after i.c.v. PACAP; including cytoskeletal, signaling and synaptic proteins |
| Guest et al. 2012 | Rt | dissection | N | LC-MS | N | Identified hypothalamic proteins that differ in expression in rats subjected to a low-protein diet as compared to wild-type controls |
| Pedroso et al. 2012 | Rt | dissection | N | MALDI-TOF MS [S]; 2D-GE, WB [V] | N | Identified 86 hypothalamic proteins in Wistar rats |
| Stelzhammer et al. 2012 | Rt | dissection | N | LC-MS/MS [S] | N | Found 21,455 peptides that corresponded to 622 unique proteins |
| X. Zhang et al. 2012 | Ms | dissection | N | LC-FT-MS/MS, SIEVE™ software-based analysis, spectral analysis | N | Identified 367 peptides from neuropeptide precursors |
| Ihnatko et al. 2013 | Ms | dissection | N | LC-MS/MS | N | Differential up- and down-regulation of proteins in tumor-bearing mice and caloric-restricted pair fed mice |
| Iqbal et al. 2013 | Rt | dissection | N | HPLC/ESI-ion trap; HPLC/ESI-Q-TOF MS | N | Identified 198 proteins, 78 of which were common to both sets of methods; 58 unique proteins identified by Q-TOF and 62 by HPLC/ESI-ion trap. |
| Kefaloyianni et al. 2013 | Rt | dissection | N | LC-MSE and LC-MS/MS | N | KATP channels in different tissues assemble with proteins having common functions |
| Taraslia et al. 2013 | Ms | dissection | N | 2D-GE, MALDI-TOF MS [S] | N | 515 different single-gene products were identified, eight of which were unique to hypothalamus |
| Iqbal et al. 2014a | Rt | dissection | N | HPLC/ESI-TOF & HPLC-Q-TOF | N | 35 and 97 significantly differentially expressed proteins by two analyses in simulated microgravity model |
| Iqbal et al. 2014b | Rt | dissection | N | HPLC/ESI-TOF & HPLC-Q-TOF | N | Differential expression of 17 specific cellular defense proteins in simulated microgravity model |
| J.-H. Kim et al. 2014 | Rt | dissection | N | LC-ESI-MS/MS [S]; WB, IHC [V] | N | Following chronic partial sleep deprivation in rats for 7 d, 89 and 50 proteins were up- and down-regulated, respectively |
| Chao et al. 2015 | Rt | dissection | N | 2D-GE, LC-MS/MS [S]; WB [V] | N | Found a few proteins induced by heatstroke that had their levels normalized by cooling |
| Zhong et al. 2015 | Ms | dissection | N | ESI-LC-MS/MS | N | Found 31 overexpressed proteins in wild-type group compared to EPHX2 KO group |
| Manousopoulou et al. 2016 | Ms | dissection | N | ESI-LC-MS/MS [S]; qPCR [V] | N | Quantitative profiling yielded 9,249 protein groups, with 7,718 groups profiled with a minimum of two unique peptides each; high-fat diet or lipopolysaccharide challenge produced unique proteomic profiles |
| Azzam et al. 2017 | Ms | dissection | N | RPLC-MS/MS [S]SRM MS [V] | N | Found 39 proteins showing differences in expression in mouse models of narcolepsy |
| Pedroso et al. 2017 | Rt | dissection | N | Q-TOF MS [S]; 2D-GE [V] | N | 1,356 proteins were identified and 348 were quantified, along with 127 metabolites. Intrauterine growth restriction resulted in down-regulation of 36 proteins and 5 metabolites, and up-regulation of 21 proteins and 9 metabolites in the hypothalamus. |
| Udvari et al. 2017 | Rt | dissection | N | 2D-DIGE [S] LC/MS-MS; WB; ISH; IHC; EM [V] |
Y | Identified 26 proteins Isolated synaptosome fractions from maternal rats, 7 of which up-regulated and 19 were down-regulated. Identified a complement cascade protein by WB, ISH, IHC and EM to be present within ARH and VMH. |
| Zettergren et al. 2017 | Ms | dissection | N | MS and MS/MS; iTRAQ [S]; PRM [V] | N | Identified 2,998 proteins in hypothalamus and amygdala of neonatal male, female and androgen receptor knockout male mice; of which 173 proteins were expressed differentially in males and females. Verified expression of seven genes using targeted proteomics. |
| Cao et al. 2018 | Gs | dissection | N | iTRAQ; LC-MS/MS [S]; qRT-PCR; WB [V] | N | Found 18 proteins up-regulated and 16 down-regulated in association with conditions of periods before and during egg laying |
| Firmino et al. 2018 | Rt | NS | N | LC-MS/MS | N | Found 7,021 proteins, many of which exhibited changes in relative abundance in immune-activated rats relative to controls |
| Govindaraj et al. 2018 | Rt | dissection | N | 2D-GE; MALDI-TOF/TOF MS [S]; semi-quant RT-PCR; WB [V] | N | Found 21 protein spots differentially expressed in preoptic, whole hypothalamic, hippocampal and pituitary tissues of females exposed neonatally to estradiol |
| Nobis et al. 2018 | Ms | dissection | N | 2D-GE; HPLC; LC/Q-TOF [S]; LC-ESI-MS/MS; WB [V] | N | Identified 22 proteins that dismayed alterations in levels in the hypothalamus among three groups: activity-based anorexic, limited-food access, and ad libitum-fed |
| X. Y. Zhang et al. 2018 | Ck | dissection | N | iTRAQ; LC-MS/MS | N | Found 235 differentially expressed proteins between L-arginine-fed and control subjects |
| ARH: Arcuate hypothalamic nucleus | ||||||
| Amigó-Correig et al. 2012 | Ms | dissection | N | MALDI-TOF/TOF | N | Adult lean and high fat diet-induced obese mice orally treated with sodium tungstate had modified levels of proteins involved in cell morphology, axonal growth and tissue remodeling |
| Preoptic area | ||||||
| Govindaraj et al. 2018 | Rt | dissection | N | 2D-GE; MALDI-TOF/TOF MS [S]; semi-quant RT-PCR; WB [V] | N | Found 21 protein spots differentially expressed in preoptic, whole hypothalamic, hippocampal and pituitary tissues of females exposed neonatally to estradiol |
| PVH: Paraventricular hypothalamic nucleus | ||||||
| Romanov et al. 2014 | Rt | dissection | Y | Illumina HiSeq2000 sequencer, MALDI-TOF | Y | Profiled secretagogin neurons as a distinct CRH-releasing neuron population |
| SCH: Suprachiasmatic hypothalamic nucleus | ||||||
| Deery, Maywood et al. 2009 | Ms | NS | N | 2D-DIGE & MS | N | 13% of soluble proteins were found to be subject to circadian regulation |
| SO: Supraoptic hypothalamic nucleus | ||||||
| Goraud et al. 2007 | Rt | dissection | N | 2D-GE, MALDI-TOF MS [S]; WB, IHC [V] | N | Identified 14–3-3 proteins that are up-regulated as a consequence of chronic dehydration |
| VLPO: Ventrolateral preoptic nucleus | ||||||
| Dooley et al. 2010 | Rt | dissection | N | MALDI-TOF/TOF MS | N | Identified diaminochlorotriazine (DACT) protein adducts formed in Atrazine-exposed rats |
| VMH: Ventromedial hypothalamic nucleus | ||||||
| Mo et al. 2006 | Rt | μ-punch | N | 2D-GE [S] LC-ESI-MS/MS [V] | N1 | Identified 99 unique proteins based on data from 2D-GE experiments, which comprise a “primary proteome database” for the VMH |
| Mo et al. 2008 | Rt | μ-punch | N | RPLC-nanoESI-MS/MS | N1 | Up-regulation of 29 identified proteins with estradiol treatment |
*
Abbreviations: 2D-DIGE, two-dimensional difference gel electrophoresis; 2D-GE, two-dimensional gel electrophoresis; 2D-LC, two-dimensional liquid chromatography; μ-punch, micropunch; cICAT, cleavable isotope-coded affinity tags; Ck, chicken; CRH, corticotropin-releasing hormone; Cw, cow; EM, electron microscopy; ESI, electrospray ionization; Fs, fish; FT, Fourier transformation; FTICR, Fourier-transform Ion Cyclotron Resonance; Gs, goose; HCD and ETD-based MS/MS, High-energy Collisional Dissociation and Electron-transfer Dissociation-based tandem mass spectrometry; IHC, immunohistochemistry; iTRAQ, isobaric tag for relative and absolute quantitation; KO, knockout; LC, liquid chromatography; LC-MSE, liquid chromatography – label-free mass spectrometry; LTQ, Linear Trap Quadrupole; MALDI-TOF, Matrix-Assisted Laser Desorption/Ionization-Time of Flight; Ms, mouse; MS/MS, tandem mass spectrometry; MSPD, Matrix Solid-Phase Dispersion; nanoESI, nano-electrospray ionization; NS, not stated; PACAP, pituitary adenylate cyclase-activating polypeptide; PRM, parallel reaction monitoring; qRT-PCR, quantitative real-time polymerase chain reaction; Q-TOF, quadrupole time of flight; RPLC, reversed-phase liquid chromatography; SELDI-TOF, Surface-Enhanced Laser Desorption/Ionization-Time of Flight; semi-quant, semi-quantitative; SRM, Selected Reaction Monitoring; Triple Quad MS, triple quadrupole mass spectrometry; Rt, rat; WB, Western blotting
1
The authors reference specific atlas plates and locations for their micropunches.