Angiotensinergic and noradrenergic mechanisms in the hypothalamic paraventricular nucleus participate in the drinking response induced by activation of the subfornical organ in rats.

The present study was done to investigate the contribution of the hypothalamic paraventricular nucleus (PVN) to the drinking response caused by activation of the subfornical organ (SFO) following angiotensin II (ANG II) injections in the awake rat. Microinjection of ANG II into the SFO elicited the drinking response. Previous injections of either saralasin, an ANG II antagonist, or phentolamine, an alpha-adrenoceptor antagonist, bilaterally into the PVN resulted in the significant attenuation of the drinking response to ANG II. Similar injections of any of the beta-adrenoceptor antagonist timolol, the muscarinic antagonist atropine, or saline vehicle into the PVN had no significant effect on the drinking response. In an attempt to clarify the neural mechanisms in the PVN involved in the drinking response to ANG II injected into the SFO, the effect of microinjection of ANG II into the SFO on noradrenaline (NA) release in the PVN was examined using intracerebral microdialysis techniques. The injection of the ANG II, but not saline vehicle, significantly enhanced the NA release in the region of the PVN. These results indicate the involvement of both the angiotensinergic and alpha-adrenergic systems in the PVN in the drinking response caused by angiotensinergic activation of the SFO, and imply that the angiotensinergic projections from the SFO to the PVN may serve to increase NA release which results in mediating water intake.

GABAergic modulation of neurons in the nucleus of the solitary tract with ascending projections to the subfornical organ in the rat.

Twenty-five neurons in the region of the nucleus of the solitary tract (NTS) were antidromically activated by electrical stimulation of the subfornical organ (SFO) in male rats under urethane anesthesia. Microiontophoretically applied bicuculline, a gamma-aminobutyric acid (GABA)(A) antagonist, but not phaclofen, a GABA(B) antagonist, attenuated the post-antidromic inhibitory response evoked by SFO stimulation of approximately two-third (n=17) of identified neurons, indicating the existence of recurrent inhibitory systems through GABA(A) receptors. Iontophoretically applied GABA decreased the spontaneous activity of all identified neurons, and the GABA-induced inhibition was prevented by simultaneously applied bicuculline, but not by phaclofen. Activation of peripheral baroreceptors, achieved by rising arterial blood pressure with an intravenous infusions of phenylepherine, suppressed the activity of the majority (n=20) of identified neurons. The inhibitory response of identified neurons (n=7) to baroreceptor activation was partially antagonized by iontophoretically applied bicuculline, but not by phaclofen. These results imply that GABAergic mechanisms may modulate the baroreceptor reflex acting on GABA(A) receptors of NTS neurons with ascending projections to the SFO in the region of the NTS.

Estrogen decreases the responsiveness of subfornical organ neurons projecting to the hypothalamic paraventricular nucleus to angiotensin II in female rats.

Extracellular single-unit activity was recorded from subfornical organ (SFO) neurons antidromically identified as projecting to the hypothalamic paraventricular nucleus (PVN) in urethane-anesthetized ovariectomized female rats that were treated with either propylene glycol (PG) vehicle or estradiol benzoate (EB). No significant differences were observed between the PG- and EB-treated rats in the latency, conduction velocity, or threshold of antidromic activation. The mean spontaneous firing rate was significantly lower and the refractory period was significantly longer in the EB-treated rats. In the identified units that were activated by angiotensin II (ANG II) applied iontophoretically, the amount of excitatory response to intracarotid administration of ANG II was much greater in the PG-treated than in the EB-treated rats. These results suggest that estrogen may decrease the responsiveness of SFO neurons projecting to the PVN to circulating ANG II.

Hemorrhage activates catecholaminergic neurons sensitive to GABA in the nucleus of the solitary tract with ascending projections to the subfornical organ in rats.

The activity of neurons in the region of the nucleus of the solitary tract (NTS) that were antidromically identified by electrical stimulation of the rat subfornical organ (SFO) was tested for a response to microiontophoretic application of gamma-aminobutyric acid (GABA), hemorrhage (10 ml/kg b.w.t.), or local administration of the chemical neurotoxin, 6-hydroxydopamine (6-OHDA), into the SFO stimulation site. Microiontophoretically (MIPh) applied GABA caused a decrease excitability in 22 out of 24 neurons tested, and the inhibition was blocked by MIPh-applied bicuculline, a GABAA antagonist, but not by phaclofen, a GABAB antagonist. Of these neurons that responded to GABA, 17 displayed an increase in neural firing in response to hemorrhage, while 5 were unresponsive. The occurrence of both antidromic spikes and post-stimulus inhibition of 9 out of 13 neurons tested was completely abolished by the injection of 6-OHDA into the SFO. These results suggest that neurons in the region of the NTS, which carry peripheral baroreceptor information to the SFO, receive GABAergic inhibitory inputs via a GABAA receptor mechanism, and imply that part of these neurons are catecholaminergic.

Retinal vascular pattern in familial exudative vitreoretinopathy.

We evaluated the retinal vascular pattern in 133 eyes (77 cases) with familial exudative vitreoretinopathy (FEVR). A wide variety of retinal vascular abnormalities were observed in the extreme fundus periphery, particularly in the temporal retinal sector. They included: presence of an avascular zone in the extreme periphery, vasodilation and arteriovenous anastomosis in the peripheral vascularized retina, v-shaped avascular zone along the temporal meridian, and neovascularization. A grading system into five types was proposed with these features as parameters. Macular ectopia, falciform retinal fold and vitreoretinal adhesion appeared to be secondary to retinal vascular abnormalities. The presence of sporadic FEVR is postulated.

Estrogen decreases the responsiveness of subfornical organ neurons to angiotensinergic neural inputs from the lateral hypothalamic area in the female rat.

Twenty-eight subfornical organ (SFO) neurons in ovariectomized (OVX) female rats that were treated with propylene glycol (PG) vehicle and 26 SFO neurons in OVX female rats that were treated with estrogen benzoate (EB) were antidromically activated by electrical stimulation of the hypothalamic paraventricular nucleus (PVN) under urethane anesthesia. No significant differences were observed between the PG-treated and EB-treated OVX animals in the latency, conduction velocity, or threshold of antidromic activation. The mean spontaneous discharge rate was significantly lower in the EB-treated than in the PG-treated OVX animals. In both groups, the activity of the majority (86% in the PG-treated animals and 88% in the EB-treated animals) of identified SFO neurons were activated by microiontophoretic application of angiotensin II (ANG II). Electrical stimulation of the lateral hypothalamic area (LHA) increased the excitability of these ANG II-sensitive SFO neurons (58% in the PG-treated animals and 52% in the EB-treated animals). The excitatory response to either ANG II or LHA stimulation was blocked by microiontophoretic application of the ANG II antagonist saralasin (Sar), suggesting that the excitatory response to LHA stimulation may be mediated by angiotensinergic LHA projections to the SFO. The magnitude of excitatory response to either ANG II or the LHA stimulation was much greater in the PG-treated than in the EB-treated animals. These results suggest that estrogen decreases the responsiveness of SFO neurons projecting to the PVN to angiotensinergic inputs from the LHA.

Retinal involvement in familial exudative vitreoretinopathy.

34 cases of familial exudative vitreoretinopathy were seen during a 40-month period ending April 1981. There were 30 cases from 11 families and 4 sporadic cases. The familial cases showed the mode of inheritance compatible with the autosomal-dominant inheritance. Ophthalmoscopy and super-wide panoramic fluorescein angiography revealed characteristic retinal vascular abnormalities in all cases, embracing supernumerous vascular branchings, arteriovenous shunt formation in the extreme periphery, generalized hyperpermeability of vessels and incomplete arteriovenous interdigitating pattern. Vitreoretinal adhesion was present in the peripheral temporal retina in all cases. An avascular zone wider than 1 disc diameter was noted in the majority of cases in the extreme temporal periphery. A V-shaped retinal degeneration was seen along the temporal meridian in 18 cases (48%). Retinal detachment was present in 19 eyes of 14 cases, and it was rhegmatogenous in 11 and traction-induced in 8 eyes. Retinal exudates and total posterior vitreous detachment was seen only occasionally.

Activation of serotonergic pathways from the midbrain raphe system to the subfornical organ by hemorrhage in the rat.

The role of serotonergic neural pathways from the midbrain raphe nuclei to the subfornical organ (SFO) in the central regulation of cardiovascular function and body fluid balance was investigated in adult male rats under urethane anesthesia. Eleven neurons in the dorsal raphe nucleus (DR) were antidromically activated by electrical stimulation of the SFO. Of these neurons, 6 displayed an excitatory response following hemorrhage (10 ml/kg bwt) while the remaining 5 neurons were unresponsive. Ninety-four neurons in the SFO were tested for a response to electrical stimulation of the DR or hemorrhage. Electrical stimulation of the DR caused orthodromic excitation (19%) or inhibition (5%) of the activity of SFO neurons. In 14 of 18 SFO neurons that displayed the excitation to the stimulation of the DR, hemorrhage (30 to 50 mm Hg suppression in mean arterial pressure) produced an increase of their discharge, while the stimulus was without effect in the remaining neurons responsive to the stimulation of the DR. The effects of hemorrhage on serotonin (5-HT) release in the region of the SFO were examined using intracerebral microdialysis techniques. Hemorrhage significantly increased 5-HT and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) concentrations in the region of the SFO. The present data suggest that the serotonergic pathways from the DR to the SFO may relay activation of the peripheral baroreceptors to SFO neurons which result in enhanced excitability, indicating the involvement of the pathways in the regulation of cardiovascular function.

Phage display of xylan-binding module of xylanase J from alkaliphilic Bacillus sp. strain 41M-1.

Xylanase J from alkaliphilic Bacillus sp. strain 41M-1 has a family 11/G catalytic domain and a xylan-binding module (XBM). The XBM of xylanase J was displayed on the surface of filamentous bacteriophage. The XBM expressed on the phage surface retained binding activity to xylan. Random mutations were introduced in the XBM gene by error-prone PCR, and the repertoire was cloned for display on phage. Sequence analysis of the xylan-binding activity-deficient mutants revealed that Phe 284 and Trp317 of the XBM would contribute to the xylan-binding activity.