Involvement of GABAergic projection from central amygdaloid nucleus to paraventricular nucleus of hypothalamus in electroacupuncture induced regulation of gastric function in GAD2-Cre mice / 针刺研究

OBJECTIVE: To explore the effect of γ-aminobutyric acid (GABA)ergic neuronal circuit of the central amygdaloid nucleus (CeA) and the paraventricular nucleus of hypothalamus (PVN) on electroacupuncture (EA)-induced regulation of gastric function by way of CeA-PVN projection. METHODS: The present study included 3 parts: 1) C57BL/6 mice were randomly divided into control and EA groups (n=6 in each group). EA was applied to right "Weishu"(BL21, Back-shu point) and "Zhongwan"(CV12, Front-mu point) for 20 min, followed by detecting the expression of c-fos in the CeA and PVN by using immunofluorescence staining; 2) Microinjection of anterograde tracer (rAAV-EF1α-DIO-mcherry-WPRE-pA) into the CeA was conducted in GAD2-Cre mice for confirming the projection of GABAergic neurons from CeA to PVN; 3) GAD2-Cre mice were randomly divided into rAAV-DIO-mcherry (intra-CeA injection of rAAV-EF1α-DIO-mcherry-WPRE-pA), rAAV-DIO-hM3D（Gq）-mcherry(intra-CeA injection of rAAV-EF1α-DIO-hM3D(Gq)-mcherry-WPRE-pA) and rAAV-DIO-hM3D（Gq）-mcherry+EA groups(n=6 in each group). The food intake and gastric empty were detected, and the concentration of GABA in the PVN was assayed by using high performance liquid chromatography on the 28th day after intra-CeA injection. RESULTS: 1) The expression of c-fos in the CeA and PVN was significantly increased in the EA group relevant to the control group(P<0.01), suggesting an activation of neurons in both CeA and PVN after EA. 2) Following CeA injection of rAAV-EF1α-DIO-mcherry-WPRE-pA, the densely expressed virus GABAergic neurons were found in CeA and large number of projection fibers found in the PVN, suggesting a direct connection between CeA and PVN. 3) After activating the GABAergic neurons of CeA, the concentration of GABA in the PVN was obviously increased (P<0.01), the food intake and the gastric empty were considerably decreased relevant to the rAAV-DIO-mcherry group(P<0.01). Following EA intervention，the concentration of GABA in the PVN was obviously decreased(P<0.01), the food intake and the gastric empty were significantly increased relevant to the rAAV-DIO-hM3D（Gq）-mcherry group (P<0.01). CONCLUSION: EA of BL21 and CV12 (Back-shu and Front-mu acupoints) can increase food intake and gastric empty in GAD2-Cre mice, which may be achieved by suppressing the release of GABA in PVN through CeA-PVN GABAergic neural circuit.

GABA in the central amygdaloid nucleus modulates the electrolyte excretion and hormonal responses to blood volume expansion in rats

We investigated the involvement of GABAergic mechanisms of the central amygdaloid nucleus (CeA) in unanesthetized rats subjected to acute isotonic or hypertonic blood volume expansion (BVE). Male Wistar rats bearing cannulas unilaterally implanted in the CeA were treated with vehicle, muscimol (0.2 nmol/0.2 µL) or bicuculline (1.6 nmol/0.2 µL) in the CeA, followed by isotonic or hypertonic BVE (0.15 or 0.3 M NaCl, 2 mL/100 g body weight over 1 min). The vehicle-treated group showed an increase in sodium excretion, urinary volume, plasma oxytocin (OT), and atrial natriuretic peptide (ANP) levels compared to control rats. Muscimol reduced the effects of BVE on sodium excretion (isotonic: 2.4 ± 0.3 vs vehicle: 4.8 ± 0.2 and hypertonic: 4.0 ± 0.7 vs vehicle: 8.7 ± 0.6 µEq·100 g-1·40 min-1); urinary volume after hypertonic BVE (83.8 ± 10 vs vehicle: 255.6 ± 16.5 µL·100 g-1·40 min-1); plasma OT levels (isotonic: 15.3 ± 0.6 vs vehicle: 19.3 ± 1 and hypertonic: 26.5 ± 2.6 vs vehicle: 48 ± 3 pg/mL), and ANP levels (isotonic: 97 ± 12.8 vs vehicle: 258.3 ± 28.1 and hypertonic: 160 ± 14.6 vs vehicle: 318 ± 16.3 pg/mL). Bicuculline reduced the effects of isotonic or hypertonic BVE on urinary volume and ANP levels compared to vehicle-treated rats. However, bicuculline enhanced the effects of hypertonic BVE on plasma OT levels. These data suggest that CeA GABAergic mechanisms are involved in the control of ANP and OT secretion, as well as in sodium and water excretion in response to isotonic or hypertonic blood volume expansion.

Role of Lateral Septum-ACh Pressor System in Central Amygdaloid Nucleus-emotional Pressor System / 医学研究杂志

Objective The present study was to analyze the role of ACh-lateral septum(SL)pressor system in the Central amygdaloid nucleus(AC)-emotional pressor circuit.Method Interurban microinjection of different drugs,then blood pressure and heart vate were vecorded.Results(1)CRF(corticotropin releasing factor)or SP(substance P)microinjection into the SL can induced pressor responses.The AC pressor responses to glutamate(Glu)could be attenuated by preinjection of CRF antagonist or SP antagonist into bilateral SL.(2)the AC pressor responses to Glu were also reduced by preinjection of atropine into either bilateral HBL,LC or RVL respectively,(3)Since lateral HB(HBL)projects to the posterior hypothalamus(HP)containing ACh-ergic neurons,and excitation of the latter produces pressor response via the RVL and LC-RVL;atropine preinjection into the HP could also decrease the HBL-and AC-pressor response.Conclusion The present results indicate that the AC produce pressor responses involved with the SL via their CRF and SP,mechanism underlying pressor response of SL is involved in pressor response of AC.

AGING CHANGES OF NEUROTENSIN-LIKE AND GALANIN-LIKE NEURONS IN THE CENTRAL AMYGDALOID NUCLEUS OF THE RAT / 解剖学报

Aging changes of neurotensin-like (NT-L) and galanin-like (GAL-L) cell bodies and fiber terminals in the central amygdaloid nucleus between the young and old rats were studied by means of immunohistochemical and microspectrophotometric techniques. NT-L cell bodies were mainly located in the central part (CeLn) of, centrolateral amygdaloid nucleus, a few of NT-L neurons were also found in the centrolateral amygdaloid nucleus, capsular part (CeLc) and centromedial amygdaloid nucleus (CeM). NT-L fiber terminals were seen mainly in the CeLn and CeLc. GAL-L cell bodies were observed only in the CeM. A marked decrease of density of NT-L cell bodies and fiber terminals in the central amygdaloid nucleus was found in the old rats as compared with young animals. No significant age difference in number of GAL-L cell bodies was observed in the central amygdaloid nucleus, but the staining intensity of GAL-L cell bodies was reduced remarkably. The results show that there are apparent changes with aging of the NT-L and GAL-L neurons in the central amygdaloid nucleus of the rat. The significance of the changes remains to be studied further.