The Effects Of REM Sleep Deprivation On Levels Of Neuropeptides In The Hypothalamus And Brainstem Of Rats

Objectives: To study the effects of sleep deprivation on thetranscription of NPB, NPW,their receptors GPR7and prepro-orexinmRNA in the hypothalamus and brainstem of rats， then To observe theeffects of sleep deprivation on levels of NPB protein, and explore therelationship between NPB, NPW, GPR7, orexin and sleep-wake cycle.Methods: Forty adult male Sprague-Dawley rats were randomly divided intocage control group，REM sleep deprivation group(ld，3d,5d),each grouphad ten rats． The sleep deprivation model was established by modifiedmultiple platforms method(MMPM).RT-qPCR was used to test thetranscription of NPB, NPW,their receptors GPR7and prepro-orexin mRNAin the hypothalamus and brainstem of rats. NPB immune positive cellswere observed in the lateral hypothalamic area and Raphe nucleus byimmunofluorescence.Result:1. The effects of REM sleep deprivation on transcriptionalLevels of NPB,NPW,their receptors GPR7and prepro-orexin mRNA inthe hypothalamus and brainstem of rats（1） In rats hypothalamus, compared with CC group， thetranscription of prepro-orexin mRNA increased obviously in RD1d and RD3dgroup (P<0.01)while the transcription of GPR7mRNA decreased (P<0.01),when it fell close to CC group, the transcription of NPB and GPR7mRNAboth began to increase (P<0.01),but RD was found have no obvious effecton NPW mRNA transcription in this study.（2）In rats midbrain, thetranscription of NPW and NPB mRNA decreased obviously in RD3d and RD5dgroup compared with CC group (P<0.01),but there is no difference betweenRD1d and CC group (P>0.05). The transcription of GPR7mRNA in RD5d groupwas significantly lower than that of CC group (P<0.01), but RD1d and RD3d group have no significant differences compared with CC group (P>0.05).The transcription of prepro-orexin mRNA increased obviously in SD1d andSD3d (P<0.01) while it decreased in SD5d but it was still higher thanthat of CC group (P<0.01).2. The effects of sleep deprivation on levels of NPB proteinImmunofluorescence showed that NPB immune positive cellsincreased in the lateral hypothalamus (LH) after SD5d while in theraphe nucleu (DR) it increased from the first day of RD,and it presenteda rising trend.3. Neuropeptide B immunofluorescence in the central nervous systemof the ratNPB immune positive (irNPB) cells were found not only prenst inthe hypothalamic(ventromedial nucleus, dorsal medial nucleus), arcuatehypothalamic nucleus,supraoptic nucleus and zona incerta, the ventraltegmental area,dorsal raphe nucleus, substantia nigra and so on of themidbrain,they also were found in the basal forebrain (VDB and HDB), globuspallidus （lateral,medial）,reticular thalamic nucleus,median raphenucleus,paramedian raphe nucleus,raphe obscurus nucleus,anteriortegmental nucleus,rhabdoid nucleus,paramedian reticular nucleus andgigantocellular reticular nucleus.Conclusions: NPB and GPR7may be involved in the regulation of sleep-wakein the hypothalamus and midbrain of rats, and the effects may becontrary to the orexin.In rats midbrain, NPW and NPB may have similareffects in the regulation of sleep-wake.Moreover,NPB may be participatedin regulation of energy metabolism and mood disorders which cased by REMsleep deprivation.