Knockdown of CXCL14 disrupts neurovascular patterning during ocular development.

The C-X-C motif ligand 14 (CXCL14) is a recently discovered chemokine that is highly conserved in vertebrates and expressed in various embryonic and adult tissues. CXCL14 signaling has been implicated to function as an antiangiogenic and anticancer agent in adults. However, its function during development is unknown. We previously identified novel expression of CXCL14 mRNA in various ocular tissues during development. Here, we show that CXCL14 protein is expressed in the anterior eye at a critical time during neurovascular development and in the retina during neurogenesis. We report that RCAS-mediated knockdown of CXCL14 causes severe neural defects in the eye including precocious and excessive innervation of the cornea and iris. Absence of CXCL14 results in the malformation of the neural retina and misprojection of the retinal ganglion neurons. The ocular neural defects may be due to loss of CXCL12 modulation since recombinant CXCL14 diminishes CXCL12-induced axon growth in vitro. Furthermore, we show that knockdown of CXCL14 causes neovascularization of the cornea. Altogether, our results show for the first time that CXCL14 plays a critical role in modulating neurogenesis and inhibiting ectopic vascularization of the cornea during ocular development.

The oestrogenized rat myometrium inhibits organotypic sympathetic reinnervation.

Chronic administration of oestrogen to rats during the infantile/prepubertal period provokes, at 28 days of age, complete loss of noradrenaline-labelled intrauterine sympathetic nerves. It is not known whether oestrogen inhibits the growth or causes the degeneration of developing uterine sympathetic nerves, or whether the uterus recovers its innervation following cessation of infantile/prepubertal oestrogen treatment. In the present study, we analysed the time-course of the effects of oestrogen on the development of uterine sympathetic nerves in the rat, using histochemical methods. In addition, the pattern of sympathetic reinnervation of the uterus of intact and ovariectomised females was assessed 3 and 6 months after cessation of chronic oestrogen treatment. The ability of sympathetic nerves to reinnervate the oestrogenized uterine tissue was assessed in intraocular transplants of uterine myometrium into ovariectomised host rats. Early exposure to oestrogen did not inhibit the approach of sympathetic nerves to the uterus, but prevented the normal growth and maturation of intrauterine sympathetic fibres and abolished the innervation that reached the organ before initiation of treatment. Three or six months following cessation of oestrogen treatment, most of the sympathetic nerves were restricted to the mesometrium and mesometrial entrance, whereas intrauterine innervation remained persistently depressed as a consequence of a sustained oestrous-like state provoked by ovarian dysfunction (polycystic ovary). An organotypic regrowth of uterine sympathetic nerves was observed in ovariectomised infantile/prepubertal oestrogen-treated animals. After 5 weeks in oculo, the innervation of oestrogenized myometrial transplants was reduced by 50%, and substantial changes in the pattern of reinnervation were observed. In control transplants, 86% of the nerves were terminal varicose myometrial and perivascular nerve fibres, whereas 14% were preterminal nerve bundles. In oestrogenized myometrial transplants, 83% of the noradrenaline-labelled intercepting nerves were enlarged preterminal bundles and only 17% were terminal fibres. These results indicate that the oestrogenized myometrium is unattractive for sympathetic nerves and inhibits organotypic sympathetic reinnervation.

Effects of chronic oestrogen treatment are not selective for uterine noradrenaline-containing sympathetic nerves: a transplantation study.

Previous studies have shown that chronic administration of oestrogen during postnatal rat development dramatically reduces the total content of noradrenaline in the uterine horn, abolishes myometrial noradrenergic innervation and reduces noradrenaline-fluorescence intensity of intrauterine perivascular nerve fibres. In the present study we analysed if this response is due to a direct and selective effect of oestrogen on the uterine noradrenaline-containing sympathetic nerves, using the in oculo transplantation method. Small pieces of myometrium from prepubertal rats were transplanted into the anterior eye chamber of adult ovariectomised host rats. The effect of systemic chronic oestrogen treatment on the reinnervation of the transplants by noradrenaline-containing sympathetic fibres from the superior cervical ganglion was analysed on cryostat tissue sections processed by the glyoxylic acid technique. In addition, the innervation of the host iris was assessed histochemically and biochemically. The histology of the transplants and irises was examined in toluidine blue-stained semithin sections. These studies showed that after 5 wk in oculo, the overall size of the oestrogen-treated transplants was substantially larger than controls, and histology showed that this change was related to an increase in the size and number of smooth muscle cells within the transplant. Chronic oestrogen treatment did not provoke trophic changes in the irideal muscle. Histochemistry showed that control transplants had a rich noradrenergic innervation, associated with both myometrium and blood vessels. Conversely, in oestrogen-treated transplants only occasional fibres were recognised, showing a reduced NA fluorescence intensity. No changes in the pattern and density of innervation or in the total content of noradrenaline of the host irises were detected after chronic exposure to oestrogen. We interpreted these results to indicate that the effects of oestrogen on uterine noradrenaline-containing sympathetic nerves are neither selective or direct, but result from an interaction between sympathetic nerve fibres with the oestradiol-primed uterine tissue. A potential effect of oestrogen on the neurotrophic capacity of the uterus is discussed.

Estudo do ciclo pupilar e de algumas manifestaçöes disautonômicas em portadores de hanseníase / Study of the pupil cycle time and some manifestations of autonomic involvement in leprosy patients

Em nosso estudo, realizou-se a avaliaçäo funcional da inervaçäo autonômica da íris de portadores das várias formas de manifestaçäo clínica da hanseníase através da medida do tempo do ciclo pupilar. Concomitantemente, procurou-se pesquisar a funçäo lacrimal, o comportamento da pressäo arterial diante das variaçöes posturais, as alteraçöes da temperatura corporal e a ocorrência de sintomas relacionados com o comprometimento do sistema nervoso autônomo. Foram incluídos 81 participantes, divididos em dois grupos: o Grupo I, composto por 63 portadores de hanseníase, e estratificado em cinco subgrupos (HT, HB, HBB, HBL e HL), dependendo da forma de manifestaçäo clínica da doença, e o Grupo II ou grupo controle, composto por 18 indivíduos hígidos. Limitamos a idade de todos os participantes entre 16 e 45 anos. Os portadores de hanseníase estavam em tratamento regular, tinham o tempo de diagnóstico inferior a 3 anos, grau de incapacidade 0 ou 1 e näo apresentavam surto reacional no momento da avaliaçäo. A medida do tempo do ciclo pupilar, realizada segundo a padronizaçäo de Miller & Thompson (1978), demonstrou ser um método ambulatorial näo invasivo, simples e objetivo para a avaliaçäo funcional da inervaçäo autonômica da íris. O tempo do ciclo pupilar observado nos integrantes dos Subgrupos HT e HBT foi estatisticamente semelhante ao dos indivíduos hígidos enquanto o tempo do ciclo pupilar dos integrantes dos Subgrupos HBB, HBL e HL apresentou um prolongamento significativo. Dentre os outros sinais disautonômicos pesquisados, apenas a frequência de disfunçäo lacrimal nos portadores das formas HBT e HL foi significativamente maior em relaçäo à dos indivíduos hígidos. Näo se verificou a ocorrência de hipotensäo arterial ortostática e nem de alteraçöes da temperatura corporal nos portadores de hanseníase. No entanto, a frequência de alguns sintomas disautonômicos foi significativamente aumentada no Grupo I em relaçäo ao Grupo II; esses sintomas foram mais frequentes no Subgrupo HL e menos no Subgrupo HT.

Parasympathetic denervation of the iris in Chagas' disease.

The parasympathetic innervation of the iris along with cardiovascular reflexes involving parasympathetic and sympathetic function were studied in 45 patients with Chagas's disease and in 36 controls. The autonomic features in Chagas' disease included 15 with cardiomyopathy, three with megaoesophagus and five with megacolon. None of the patients had orthostatic hypotension. Parasympathetic cardiac reflexes (deep breathing 30:15 ratio, Valsalva) were abnormal. There were exaggerated pupillary responses to dilute pilocarpine. These studies suggest iris parasympathetic denervation and favour more widespread cholinergic involvement in patients with Chagas' disease, than previously recognized.

Morphine-induced pupillary fluctuation: physiological evidence against selective action on the Edinger-Westphal nucleus.

In the rat, the predominant pupillary effect of morphine sulfate (30 mg/kg, i.p.) is mydriasis, interrupted periodically by miotic excursions. Using infrared video pupillometry, respirometry and EEG recording, we had previously reported that miotic excursions are always correlated with the onset of EEG bursting, while mydriatic excursions are preceded by respiratory slowing and correlated with EEG burst cessation. In the current study, we found that during morphine-induced EEG bursting and related miosis, delivery of an alerting stimulus to the rat caused an immediate conversion sequence composed of respiratory slowing, burst cessation and mydriatic excursion. We also simulated morphine-induced EEG bursting through non-opioid means by delivering kindling electrical stimuli to the amygdala. When the stimulus was given during morphine-induced mydriasis, the stimulus-induced bursting resulted in a miotic excursion identical to spontaneous morphine-induced miotic excursions. These results indicate 1) that the reticular activating system is involved in morphine-induced mydriatic excursions and 2) that EEG bursting is not simply correlated with morphine-induced miotic excursions, but causes them. Collectively, these results strengthen our hypothesis that the Edinger-Westphal nucleus is not the site in which the selective, receptor-mediated pupillary effects of morphine are initiated.