TY - JOUR

T1 - Pre-treatment of blood samples reveal normal blood hypocretin/orexin signal in narcolepsy type I

AU - Aegidius, Helene M.

AU - Kruse, Lars

AU - Christensen, Gitte L.

AU - Lorentzen, Marc P.

AU - Jorgensen, Niklas R.

AU - Moresco, Monica

AU - Pizza, Fabio

AU - Plazzi, Giuseppe

AU - Jennum, Poul J.

AU - Kornum, Birgitte R.

PY - 2021

Y1 - 2021

N2 - The hypocretin/orexin system regulates arousal through central nervous system mechanisms and plays an important role in sleep, wakefulness and energy homeostasis. It is unclear whether hypocretin peptides are also present in blood due to difficulties in measuring reliable and reproducible levels of the peptides in blood samples. Lack of hypocretin signalling causes the sleep disorder narcolepsy type 1, and low concentration of cerebrospinal fluid hypocretin-l/oreadn-A peptide is a hallmark of the disease. This measurement has high diagnostic value, but performing a lumbar puncture is not without discomfort and possible complications for the patient. A blood-based test to assess hypocretin-1 deficiency would therefore be of obvious benefit. We here demonstrate that heating plasma or scrum samples to 65 degrees C for 30 min at pH 8 significantly increases hypocretin-1 immunoreactivity enabling stable and reproducible measurement of hypocretin-1 in blood samples. Specificity of the signal was verified by high-performance liquid chromatography and by measuring blood samples from mice lacking hypocretin. Unspecific background signal in the assay was high. Using our method, we show that hypocretin-1 immunoreactivity in blood samples from narcolepsy type 1 patients does not differ from the levels detected in control samples. The data presented here suggest that hypocretin-1 is present in the blood stream in the low picograms per millilitres range and that peripheral hypocretin-1 concentrations are unchanged in narcolepsy type 1.

AB - The hypocretin/orexin system regulates arousal through central nervous system mechanisms and plays an important role in sleep, wakefulness and energy homeostasis. It is unclear whether hypocretin peptides are also present in blood due to difficulties in measuring reliable and reproducible levels of the peptides in blood samples. Lack of hypocretin signalling causes the sleep disorder narcolepsy type 1, and low concentration of cerebrospinal fluid hypocretin-l/oreadn-A peptide is a hallmark of the disease. This measurement has high diagnostic value, but performing a lumbar puncture is not without discomfort and possible complications for the patient. A blood-based test to assess hypocretin-1 deficiency would therefore be of obvious benefit. We here demonstrate that heating plasma or scrum samples to 65 degrees C for 30 min at pH 8 significantly increases hypocretin-1 immunoreactivity enabling stable and reproducible measurement of hypocretin-1 in blood samples. Specificity of the signal was verified by high-performance liquid chromatography and by measuring blood samples from mice lacking hypocretin. Unspecific background signal in the assay was high. Using our method, we show that hypocretin-1 immunoreactivity in blood samples from narcolepsy type 1 patients does not differ from the levels detected in control samples. The data presented here suggest that hypocretin-1 is present in the blood stream in the low picograms per millilitres range and that peripheral hypocretin-1 concentrations are unchanged in narcolepsy type 1.

KW - hypocretin

KW - orexin

KW - plasma hypocretin-1

KW - narcolepsy type 1

KW - radioimmunoassay

KW - PLASMA OREXIN-A

KW - EXPRESSION

KW - RECEPTORS

KW - NEURONS

KW - NEUROPEPTIDE

KW - LEPTIN

KW - CELLS

U2 - 10.1093/braincomms/fcab050

DO - 10.1093/braincomms/fcab050

M3 - Journal article

C2 - 33977264

VL - 3

JO - Brain Communications

JF - Brain Communications

SN - 2632-1297

IS - 2

M1 - 050

ER -