Kazumasa Honda

Afferents originating from the dorsal penile nerve excite oxytocin cells in the hypothalamic paraventricular nucleus of the rat

Electrical stimulation of the dorsal penile nerve (DPN) produced orthodromic excitation in about half of oxytocin cells in the paraventricular nucleus (PVN). In contrast, less than 10% of vasopressin cells were excited. Tactile stimulation of the glans penis by a paintbrush produced excitation in 40% of oxytocin cells. Castration did not prevent activation of oxytocin cells. These results suggest that somatosensory information from the penis is transmitted to the PVN through the DPN and that such afferent input preferentially innervates oxytocin cells.

MEDIAN PREOPTIC NEURONES PROJECTING TO THE SUPRAOPTIC NUCLEUS ARE SENSITIVE TO HAEMODYNAMIC CHANGES AS WELL AS TO RISE IN PLASMA OSMOLALITY IN RATS

Extracellular single unit activity was recorded from 73 neurones in the median preoptic nucleus (MnPO), identified by antidromic activation as projecting to the supraoptic nucleus (SON) area in urethane‐anaesthetized male rats. Thirteen of 73 identified MnPO neurones were silent, and 44 of 60 spontaneously active MnPO neurones were tested for their responses to electrical stimulation of the nucleus tractus solitarius (NTS). The cells were divided into 4 groups according to their responses; those which were excited orthodromically (OD+; n = 15), those which were unresponsive (UN; n = 21), those which were inhibited orthodromically (OD−; n=4), those which showed initial inhibition followed by excitation (OD+ n = 4). Some of these neurones were further tested for their responses to haemorrhage and/or increase in blood pressure produced by intravenous administration of the oe‐agonist, phenylephrine, and/or to hyperosmotic stimulation produced by intraperitoneal injection of 1.5 M NaCl. Six out of 10 OD+ cells were excited by haemorrhage, 6 out of 11 OD+ cells were inhibited by phenylephrine, and 5 out of 9 OD+ cells were excited by hypertonic saline. On the other hand the UN cells tended to be unresponsive to each type of stimulus. Three out of 7 OD+ cells were excited by both haemorrhage and hypertonic saline, and 3 out of 8 OD+ cells were inhibited by phenylephrine and excited by hypertonic saline. The results may suggest that MnPO neurones which receive afferent input from the NTS may be sensitive not only to haemodynamic change but also to change in plasma osmotic pressure and that such population of MnPO neurones may integrate a part of the haemodynamic and osmotic information and contribute to the control of neurohypophysial hormone release.

The contribution of the median preoptic nucleus to renal sympathetic nerve activity increased by intracerebroventricular injection of hypertonic saline in the rat

The median preoptic nucleus (MnPO) of the hypothalamus is involved in the osmotic control of neurohypophysial hormone release and drinking behavior. At the same time, renal sympathetic nerves exert multiple effects on renal functions such as regulating renal blood flow and urinary sodium excretion. We made the hypothesis that the MnPO may also regulate body fluid balance by exerting an influence on renal sympathetic nerve activity (RSNA). In this study we examined the effect of electrical stimulation of the MnPO on RSNA and the contribution of the MnPO to the change of RSNA induced by intracerebroventricular injection of hypertonic saline in the male Wistar rat. Electrical stimulation of the MnPO and the paraventricular nucleus of the hypothalamus (PVN) elicited an increase in RSNA. This increase of RSNA elicited by electrical stimulation of the MnPO was reduced by microinjections (100 nl) of 10% lidocaine or 4 mM cobaltous chloride (a synaptic transmission blocking agent) bilaterally into the PVN. Both RSNA and the mean arterial pressure (MAP) were increased by the injection of 1.5 M NaCl into the third ventricle, although heart rate (HR) was not significantly changed. These responses of RSNA and MAP were diminished by microinjection of 10% lidocaine (100 nl) into the MnPO. Our results suggest that the MnPO is involved in body fluid regulation not only by controlling vasopressin secretion and water intake but also by modulating central sympathetic outflow which regulates body fluid balance through an effect on the kidney.

Variation in the Expression of Orexin and Orexin Receptors in the Rat Hypothalamus During the Estrous Cycle, Pregnancy, Parturition, and Lactation

The widespread distribution of mRNA encoding orexin-1 (OX1R) and -2 receptors (OX2R) in the central nervous system suggests that orexin may be involved in multiple functional pathways. Central administration of orexin stimulates feeding and also affects ovarian steroid-dependent luteinizing hormone secretion, suggesting involvement of orexin in the regulation of reproductive function. To investigate a possible role for orexin in reproductive function, we examined variations in prepro-OX, OX1R, and OX2R mRNA levels in the female rat hypothalamus during the estrous cycle, pregnancy, parturition, and lactation using competitive reverse transcription-polymerase chain reaction. During the estrous cycle, only OX1R mRNA expression during late proestrus was significantly higher than that at metestrus. The prepro-OX and OX1R mRNA levels on d 1 of lactation were significantly higher than that during late pregnancy and lactation. Immunohistochemistry revealed the presence of orexin-A immunoreactive cells and the OX1R subtype in the lateral hypothalamic area as well as the magnocellular neurons of the paraventricular (PVN) and supraoptic (SON) nuclei, respectively, in pregnant and lactating rats. These results suggest a role for orexin in reproduction that may be involved in regulating physiological function in early lactation through important binding sites in hypothalamic PVN and SON.

Excitation of oxytocin cells in the hypothalamic supraoptic nucleus by electrical stimulation of the dorsal penile nerve and tactile stimulation of the penis in the rat.

In urethane-anaesthetized male rats, electrical stimulation of the dorsal penile nerve (DPN) excited 29 of 48 (60%) oxytocin cells in the contralateral supraoptic nucleus, whereas only 5 of 28 (18%) vasopressin cells were excited by the stimulation. The stimulus applied to the ipsilateral DPN to the recorded neurone also excited a similar proportion of oxytocin cells (25 of 43; 58%). Tactile stimulation of the glans penis excited 7 of 12 (58%) oxytocin cells, whereas the same stimulation excited only 3 of 16 (19%) vasopressin cells. The results suggest that sensory information arising from the penis preferentially excites oxytocin cells in the supraoptic nucleus.

Activation of paraventricular neurosecretory cells by local osmotic stimulation of the median preoptic nucleus

Both electrical and local osmotic stimulation of the median preoptic nucleus (MnPO) predominantly produced excitation of paraventricular (PVN) neurosecretory cells in the rat. By contrast osmotic stimulation of the medial septal region was without effect, although electrical stimulation excited most cells. The results suggest that the MnPO is one of the osmosensitive sites controlling electrical activity of PVN neurosecretory cells.

Electrophysiological evidence for neural connections between the supraoptic nuclei.

Effects of electrical stimulation of the contralateral supraoptic nucleus (SON) on the activity of neurosecretory neurons in the SON were studied in urethane-anesthetized lactating rats. Thirty-one out of 41 oxytocin neurons were excited and only two neurons were inhibited by contralateral SON stimulation. Eight out of 26 vasopressin neurons were excited and 8 were inhibited. These responses were not affected by suckling stimuli. Neither oxytocin nor vasopressin neurons tested were antidromically activated by contralateral SON stimulation. Thus monosynaptic connections between the bilateral SON neurosecretory neurons seem to be very few, if any. These results suggest that neural connections exist between the bilateral SON and that they are mainly polysynaptic.

Leptin affects the electrical activity of neurones in the hypothalamic supraoptic nucleus

The present experiments were undertaken to examine whether leptin affects the electrical activity of neurones in the supraoptic nucleus (SON) by using brain slice preparation of male Wistar and obese Zucker rats. Bath application of leptin (10(-8) - 10(-12) M) induced mainly inhibitory response in SON neurones of Wistar rats, although a minority showed excitation. These effects were observed in both continuously and phasically active cells. The inhibitory effect of leptin still persisted in low Ca(2+), high Mg(2+) medium. Bath application of tolbutamide, which is known to inhibit ATP-sensitive potassium channel activity, did not reverse the inhibitory effect of leptin on SON neurones. The effect of bath application of leptin was also tested in SON neurones of obese Zucker rats. Although leptin still affected the electrical activity of some SON neurones of Zucker rats, the proportion of unaffected neurones was significantly higher than in Wistar rats. The results suggest that leptin may inhibit the secretion of both oxytocin and vasopressin by inhibiting the electrical activity of neurones in the SON via direct action. This inhibitory effect of leptin may be exerted through mechanisms other than activation of ATP-sensitive potassium channels.

MILK EJECTION BURSTS OF SUPRAOPTIC OXYTOCIN NEURONES DURING BILATERAL AND UNILATERAL SUCKLING IN THE RAT

Extracellular recordings of the electrical activity of oxytocin neurones were made from the supraoptic nuclei (SON) of lactating rats, and the milk‐ejection bursts and the background activity of oxytocin neurones were investigated during unilateral and bilateral suckling. When application of pups was limited to the nipples on either the same side (ipsilateral suckling) or the side opposite (contralateral suckling) to the oxytocin neurone recorded, the burst amplitude and background firing rate were significantly (P<0.05) lower and the inter‐burst interval was significantly (P<0.05) longer than during bilateral suckling. Furthermore, the burst amplitude was significantly (P<0.05) lower during ipsilateral suckling than during contralateral suckling. The majority of the oxytocin neurones showed a gradual increase in the burst amplitude during bilateral (88.9%) and contralateral (77.3%) suckling, but during ipsilateral suckling only 40% of the neurones did. The inter‐burst interval became shorter with the progress of the milk ejection reflex during any mode of suckling. Three pairs of oxytocin neurones recorded simultaneously from both SON were successfully tested for the effect of bilateral and unilateral suckling on the electrical activity, and the results showed the same direction of change in the burst amplitude, background activity and burst interval as shown in single side recordings. These findings indicate that the burst amplitude mainly depends on the amount of afferent suckling signals arising from the nipples on the side opposite to the recording side, and that there may exist bilateral summation centres coordinating with the synchronization mechanism of milk‐ejection bursts of oxytocin neurones.

Activation of supraoptic neurosecretory cells by local osmotic stimulation of the median preoptic nucleus

In urethane-anesthetized male rats extracellular recordings were obtained from neurosecretory cells in the supraoptic nucleus (SON) while each of electrical stimulation and local osmotic stimulation produced by pressure injection of hypertonic saline was applied to the median preoptic nucleus (MnPO) or to the medial septal nucleus (MS). Electrical stimulation of the MnPO produced orthodromic excitation or initial inhibition followed by strong excitation in most SON cells (59/61), and local osmotic stimulation of the MnPO excited majority of cells (39/51). On the contrary, local osmotic stimulation of the MS did not excite SON cells, although electrical stimulation excited all the cells (5/5). The results suggest that the MnPO is one of the osmosensitive sites controlling electrical activity of SON neurosecretory cells.