Activation of hypothalamic RIP‐Cre neurons promotes beiging of WAT via sympathetic nervous system

Activation of brown adipose tissue (BAT) and beige fat by cold increases energy expenditure. Although their activation is known to be differentially regulated in part by hypothalamus, the underlying neural pathways and populations remain poorly characterized. Here, we show that activation of rat‐insulin‐promoter‐Cre (RIP‐Cre) neurons in ventromedial hypothalamus (VMH) preferentially promotes recruitment of beige fat via a selective control of sympathetic nervous system (SNS) outflow to subcutaneous white adipose tissue (sWAT), but has no effect on BAT. Genetic ablation of APPL2 in RIP‐Cre neurons diminishes beiging in sWAT without affecting BAT, leading to cold intolerance and obesity in mice. Such defects are reversed by activation of RIP‐Cre neurons, inactivation of VMH AMPK, or treatment with a β3‐adrenergic receptor agonist. Hypothalamic APPL2 enhances neuronal activation in VMH RIP‐Cre neurons and raphe pallidus, thereby eliciting SNS outflow to sWAT and subsequent beiging. These data suggest that beige fat can be selectively activated by VMH RIP‐Cre neurons, in which the APPL2–AMPK signaling axis is crucial for this defending mechanism to cold and obesity.     

兔脊髓α受体对血量扩张引起肾交感神经活动抑制和促钠排泄的作用

在窦主动脉去神经麻醉兔观察阻断脊髓α受体对血量扩张引起肾交感神经活动（RSNA）抑制和促钠排泄反应的影响。兔脊髓蛛网膜下腔注射a肾上腺素能受体阻断剂酚妥拉明与人工脑脊液后,血量扩张引起RSNA抑制分别为（－25．4±5．4）％与（－42．5±5.2）％（P＜0.05）;兔脊髓蛛网膜下腔注射α1受体阻断剂哌唑嗪与人工脑脊液后血量扩张引起RSNA抑制分别为（－29．3±6.1）％与（－42．5±5.2）％（P＜005）。结果表明,阻断脊髓α受体或α1受体均可减弱血量扩张引起RSNA抑制。脊髓注射哌唑嗪后血量扩张引起促钠排泄与利尿反应也显著减弱（P＜005）。     

Anterior neural centres in echinoderm bipinnaria and auricularia larvae: cell types and organization

Serial and interval electron micrograph series were used to examine the anterior part of the ciliary band system in the bipinnaria larva of Pisaster ochraceus and the auricularia larva of Stichopus californicus for evidence of ganglion‐like organization. The bipinnaria has paired concentrations of Multipolar with Apical Processes (MAP) cells in this region that correspond in position with previously identified clusters of serotonergic and peptidergic neurones. MAP cells located in the centre of the band have well‐developed apical processes, but no cilium. Those at the sides of the band have fewer processes, but some have recumbent cilia that extend under the glycocalyx, suggesting a sensory function. Comparable cell types are not found elsewhere in the band, a clear indication that the apical parts of the ciliary band system are organized in a distinctive fashion. Two neuronal cell types were identified in the apical region of the auricularia larva, a conventional bipolar neurone that corresponds with previously described serotonergic apical cells, and more numerous MAP cells for which there is no previous record and hence, no known transmitter. Previous immunocytochemical studies are summarized and re‐examined in the light of these results. Relevant evolutionary issues are also discussed, but the data fail to provide strong evidence either for or against Garstang’s hypothesis that the chordate brain and spinal cord derive from larval ciliary bands resembling those of modern echinoderms.     

A comparative study of the adrenergic nerves to the anterior eye segment of some primates

Summary The distribution of adrenergic terminals to the anterior eye segment of humans, Cynomolgue monkeys, squirrel monkeys, owl monkeys, Cebus monkeys, vervets, tamarins, and baboons has been investigated. The cornea is normally devoid of adrenergic terminals, except in a plexus near the limbus. The trabecular meshwork contains varying numbers of adrenergic terminals: usually none in Cynomolgus monkeys, patas monkeys, vervets, and humans, although fibres have very rarely been observed in Cynomolgus monkeys, vervets, and humans; a few in owl monkeys, squirrel monkeys, and tamarins; and moderate numbers in Cebus monkeys and baboons. From the evidence, however, it seems premature to presume an adrenergic innervation of the trabecular mechanism regulating the outflow resistance. The dilatator pupillae is regularly supplied with numerous adrenergic terminals and in the iris stroma there is probably an adrenergic innervation of the melanophores. The sphincter pupillae regularly contains adrenergic terminals with notable species differences; most fibres occur in baboons and fewest in humans, with the remaining species forming a middle class. The ciliary processes in all species contain a moderate number of adrenergic terminals, presumably primarily associated with the epithelium. Intraepithelial adrenergic terminals have been observed on the pars plana of the ciliary body of humans, Cebus monkeys, vervets, baboons, and patas monkeys. The ciliary muscle of baboons and Cynomolgus monkeys contains numerous adrenergic terminals. Moderate numbers occur in Cebus monkeys and vervets, and still less in (in falling order) tamarins, squirrel monkeys, humans, and patas monkeys.     

Fluorescent histochemical studies on the effects of 6-hydroxydopamine on adrenaline-containing nerves in the toad

Summary Fluorescence histochemistry has been used to study the effects of 6-hydroxydopamine (6-OHDA) (100 mg/kg injected into the dorsal lymph sac) on adrenaline-containing nerves in the large intestine, mesentery, lung, bladder and heart atria of the toad Bufo marinus. A gradual decrease both in fluorescence intensity and in number of detectable fibres during the first 4 hours after 6-OHDA was accompanied by a build-up of fluorescence in the nonterminal regions. These phenomena have been discussed in relation to the time course of the degeneration produced by 6-OHDA in noradrenergic nerves of higher vertebrates. Almost complete chemical sympathectomy was seen after one day, and it was not till 13 days that regenerating nerve fibres were seen in any organ. In the large intestine, however, re-innervation was slower, being first noted after 39 days. The time course of regeneration has been compared with that following sympathectomy in various mammalian organs.     

Fine structure of the autonomic nerves of the rabbit myometrium

Summary The fine structure of the preterminal nerve fibers of the rabbit myometrial smooth muscle was studied using potassium permanganate fixation or glutaraldehyde fixation with postosmification. The preterminal fibers were mostly formed by 2–10 axons enveloped by Schwann cells. Two kinds of axons and axon terminals were found. (1) Adrenergic axons, which contained many small, granular vesicles (diameter 300–600 Å) and large granular vesicles (diameter 700–1200 Å) which represented ca. 2% of the total count of the vesicles. (2) Nonadrenergic axons, which contained small agranular vesicles (diameter 300–600 Å) and large granular vesicles (diameter 700–1200 Å). Both types of axons formed preterminal varicosities along their course. The real terminal varicosities, representing the anatomical end of the axons, were usually larger than the preterminal ones and showed close contact to the plasma membranes of the smooth muscle cells. Both adrenergic and nonadrenergic terminals were found close to the smooth muscle cells, but a gap of at least 2000 Å was always present between the two cell membranes. The axons and preterminal varicosities of both types of nerves were in intimate contact with each other within the preterminal nerve fiber. Axo-axonal interactions between the two types of axons are possible in the rabbit myometrium. The relative proportion of the nonadrenergic axons from the total was about one fourth.     

The intrinsic myenteric innervation of the hind-gut and accessory muscles of defaecation in the cat

Summary The anatomy and intrinsic innervation of the colon, rectum, internal anal sphincter, ano-coccygeus and recto-coccygeus have been studied in the cat with cholinesterase and catecholamine-fluorescence histochemical techniques. A variable pattern of intrinsic innervation by acetylcholinesterase-positive and adrenergic nerves along the length of the large bowel is described and is related to segmental variations in motor activity. A variation in the distribution of non-specific cholinesterase within the muscle layers is also described. Adrenergic nerves in proximal colon are arranged in the usual peri-ganglionic manner but there is also a rich direct adrenergic innervation of the longitudinal muscle in distal colon and rectum, and of circular muscle in lower rectum and internal anal sphincter. This distribution has not been reported in other species. Direct adrenergic innervation of muscle cells has been confirmed at ultrastructural level after treatment with 5-hydroxydopamine. Adrenergic neurones have not been detected in cat bowel. The ano- and recto-coccygeus muscles and internal anal sphincter possess a dense innervation of adrenergic and cholinesterase-positive nerves. It is suggested that the variation in intrinsic innervation along the large bowel should be considered in the interpretation of pharmacological and physiological experiments on this part of the gut.This work was supported by a grant from the King's College Hospital Voluntary Research Trust. We wish to thank Dr. J. P. Tranzer and F. Hoffman-La Roche & Co. Ltd., Basle, for the gift of 5-hydroxydopamine.We also thank Miss M. K. Egan and Mr. K. J. Davies for their technical assistance.     

Parasympathetic and sympathetic postganglionic synapses in ureterovesical autonomic pathways

Summary Histochemical techniques for acetylcholinesterase and catecholamine show that ureterovesical ganglia of both cat and dog contain dense intraganglionic cholinergic and adrenergic plexuses. Ramifications of both plexuses surround most cholinergic and adrenergic ganglion cell bodies as pericellular synaptic plexuses. Similar pericellular plexuses exist around extraganglionic cholinergic and adrenergic ganglion cells. Both adrenergic and cholinergic synaptic fibers persist in denervated pregnaglionic nerve-free specimens, indicating that cholinergic synaptic fibers are postganglionic parasympathetic in nature. The presence of adrenergic (postganglionic sympathetic) and postganglionic parasympathetic synapses around cell bodies in ureterovesical ganglia provides a morphologic basis for the sympathoinhibitory and muscarinic parasympatho-excitatory phenomena described in these ganglia.     

Dense-core vesicles in motor nerve terminals

Summary Motor nerve terminals on white and intermediate muscle fibers of the Atlantic hagfish (Myxine glutinosa, L.) contain translucent synpatic vesicles and about 1–2% dense-core vesicles. Terminals on red muscle fibers contain up to 40% dense-core vesicles with diameter 800–1100 Å. Examinations for formaldehyde-induced fluorescence indicate yellow fluorescence (5-HT ?) apparently corresponding with terminal axons on red muscle fibers in craniovelar muscles. Possibly red muscle fibers of Myxine receive monoaminergic innervation.The author is indebted to Dr. Finn Walvig, Biological station, University of Oslo, Drøbak, for supply of hagfishes.     

P2X7 receptor inhibition attenuated sympathetic nerve sprouting after myocardial infarction via the NLRP3/IL‐1β pathway

Mounting evidence supports the hypothesis that inflammation modulates sympathetic sprouting after myocardial infarction (MI). The myeloid P2X₇ signal has been shown to activate the nucleotide‐binding and oligomerization domain‐like receptor family pyrin domain‐containing 3 (NLRP3) inflammasome, a master regulator of inflammation. We investigated whether P2X₇ signal participated in the pathogenesis of sympathetic reinnervation after MI, and whether NLRP3/interleukin‐1β (IL‐1β) axis is involved in the process. We explored the relationship between P2X₇ receptor (P2X₇R) and IL‐1β in the heart tissue of lipopolysaccharide (LPS)‐primed naive rats. 3′‐O‐(4‐benzoyl) benzoyl adenosine 5′‐triphosphate (BzATP), a P2X₇R agonist, induced caspase‐1 activation and mature IL‐1β release, which was further neutralized by a NLRP3 inhibitor (16673‐34‐0). MI was induced by coronary artery ligation. Following infarction, a marked increase in P2X₇R was localized within infiltrated macrophages and observed in parallel with an up‐regulation of NLRP3 inflammasome levels and the release of IL‐1β in the left ventricle. The administration of A‐740003 (a P2X₇R antagonist) significantly prevented the NLRP3/IL‐1β increase. A‐740003 and/or Anakinra (an IL‐1 receptor antagonist) significantly reduced macrophage infiltration as well as macrophage‐based IL‐1β and NGF (nerve growth factor) production and eventually blunted sympathetic hyperinnervation, as assessed by the immunofluorescence of tyrosine hydroxylase (TH) and growth‐associated protein 43 (GAP 43). Moreover, the use of Anakinra partly attenuated sympathetic sprouting. This indicated that the effect of P2X₇ on neural remodelling was mediated at least partially by IL‐1β. The arrhythmia score of programmed electric stimulation was in accordance with the degree of sympathetic hyperinnervation. In vitro studies showed that BzATP up‐regulated secretion of nerve growth factor (NGF) in M1 macrophages via IL‐1β. Together, these data indicate that P2X₇R contributes to neural and cardiac remodelling, at least partly mediated by NLRP3/IL‐1β axis. Therapeutic interventions targeting P2X₇ signal may be a novel approach to ameliorate arrhythmia following MI.