Jiro Noguchi

Stimulation of corticotropin-releasing hormone-mediated adrenocorticotropin secretion by central administration of prolactin-releasing peptide in rats.

Prolactin-releasing peptide (PrRP) is a recently isolated hypothalamic peptide which is an endogenous ligand to an orphan receptor. We previously demonstrated that PrRP neurons are widely distributed throughout the rat brain and suggested that PrRP may have important functions in the central nervous system. To analyze the function of PrRP, we studied the effect of intracerebroventricular (i.c.v.) PrRP administration on c-Fos protein accumulation in the rat brain. The results clearly indicated that c-Fos protein accumulation was dramatically increased in the nuclei of corticotropin-releasing hormone (CRH)-positive parvocellular neurosecretory cells in the paraventricular nucleus (PVN). We also demonstrated synapse-like contact between PrRP neurons and CRH cell bodies in the PVN, which suggests that PrRP31 has some effect on CRH secretion. We therefore investigated the effect of i.c.v. administration of PrRP31 on the CRH-mediated increase in adrenocorticotropin (ACTH) levels, and found that plasma ACTH levels were indeed increased by i.c.v. PrRP31. In addition, animals pre-treated with intravenous alpha-helical CRH, a potent CRH antagonist, showed attenuated plasma ACTH responses after i.c.v. PrRP31 administration. These results strongly suggest that PrRP affects the hypothalamic-pituitary-adrenal axis.

Central administration of prolactin-releasing peptide stimulates oxytocin release in rats

The prolactin-releasing peptide (PrRP) is a novel hypothalamic peptide that has been purified as a ligand of an orphan receptor which is expressed in pituitary cells, and is known to stimulate prolactin release both in vitro and in vivo. We previously determined the immunocytochemical localization of PrRP neurons in the rat brain and our results suggest that PrRP takes part in a variety of brain functions. Additionally, in rats we have demonstrated the synaptic contact of PrRP neurons with oxytocin cell bodies in the paraventricular hypothalamic nucleus (PVH) and supraoptic nucleus (SON). This observation indicates that PrRP may regulate oxytocin secretion. In the present study, we performed intra-cerebroventricular administration of PrRP to conscious rats, and examined the effect of PrRP on the plasma levels of oxytocin and vasopressin. Our results show that central administration of PrRP increased the plasma oxytocin and vasopressin levels in female rats, but in male rats only oxytocin was increased. These results suggest that the PrRP acts as a neuro-modulator of the function of magnocellular neurons, especially oxytocin neurons, in the brain.

Effects of intracerebroventricular administration of pituitary adenylate cyclase-activating polypeptide (PACAP) on the motor activity and reserpine-induced hypothermia in murines

We investigated the effects of i.c.v. administration of pituitary adenylate cyclase-activating polypeptide (PACAP) on the spontaneous motor activity and reserpine-induced hypothermia in murines. The administration of PACAP (1 or 2 nmol) caused a dose-dependent increase in both spontaneous motor activity and rearing behavior in the rat. The peptide (0.1 or 0.2 nmol) counteracted reserpine-induced hypothermia in a dose-dependent manner in mice. On the other hand, i.c.v. injection of vasoactive intestinal polypeptide, which is structurally similar to PACAP, at a dose similar to that of PACAP (2 nmol in rats, 0.2 nmol n mice) did not show a significant effect on either behavior or body temperature. Therefore, the stimulating effect of PACAP observed here may be mediated by PACAP-specific (type I) receptors. PACAP was more potent and longer-lasting than a known potent stimulating peptide, thyrotropin-releasing hormone, in both stimulating motor activity and counteracting reserpine-induced hypothermia. Results of the present study, in combination with those of previous studies identifying endogenous PACAP in the brain, suggest that PACAP may play a important role in the CNS as a stimulant in regulating motor activity and body temperature.

A novel method for counting spontaneous motor activity in the rat.

Motor activity is a good index for studying the effects of pharmacological agents. Previous investigations have measured spontaneous motor activity by counting the number of times that an animal interrupts a magnetic field or photocell beam. Quite recently, a novel activity-monitoring system, Supermex, was developed. In this system, a sensor detects the radiated body heat of an animal. The Supermex method enables an investigator to perform multi-channel measurement at low cost. Any size home cage may be used, as long as its geometry cannot block the beams contact with the animal. Operation is very simple and sensitivity adjustment is not required after the sensor-mount position and height from the cage floor are fixed. In the present study we first used the Supermex system to examine the effects of intracerebroventricular (i.c.v.) administration of a known stimulant, thyrotropin-releasing hormone (TRH). Our results confirmed the suitability of this system for testing spontaneous motor activity. We further studied the effects of pituitary adenylate cyclase-activating polypeptide (PACAP) and its related peptide, vasoactive intestinal polypeptide (VIP), on spontaneous motor activity.

A PEGylated analog of short-length Neuromedin U with potent anorectic and anti-obesity effects

Neuromedin U (NMU) is a neuropeptide known to regulate food intake and energy homeostasis that is widely distributed in the gastrointestinal tract, hypothalamus, and pituitary. A short form of NMU, porcine NMU-8 has potent agonist activity for the receptors NMUR1 and NMUR2; however, its short half-life precludes its effective use in vivo. To address this limitation, we designed and synthesized NMU-8 analogs modified by polyethylene glycol (PEG) with a molecular weight of 30kDa (PEG30k) via a variety of linkers (i.e., ω-amino- and ω-imino-carboxylic acid linker). Integrated evaluation of NMUR1 and NMUR2 binding affinities in vitro and anorectic activity in mice revealed that the introduction of a linker with a rigid ring group, e.g., 2-(piperazin-1-yl)acetic acid (PipAc), yielded a highly potent anorectic peptide, PEG30k-PipAc-NMU-8 (14), possessing improved receptor binding affinity. Subsequent optimization of the molecular weight of the PEG moiety led to the discovery of a PEG20k conjugate (15), which exhibited significant anti-obesity effect upon once-daily subcutaneous administration in diet-induced obese mice with 10% and 22% body weight loss at doses of 10 and 30nmol/kg, respectively. In addition, 15 reduced the weights of the liver and adipose tissue in a dose-dependent manner and improved the plasma biochemical parameters, e.g., insulin, glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, and total cholesterol. Thus, our results suggest that 15 (NMU-0002), which showed potent and long-lasting biological profiles in vivo, represents a candidate peptide for investigating the central and peripheral actions of NMU and its potential for clinical use.

PEGylated neuromedin U-8 shows long-lasting anorectic activity and anti-obesity effect in mice by peripheral administration

HIGHLIGHTSIntraperitoneal injection of neuromedin U‐23 and neuromedin S inhibited food intake.PEGylation gave neuromedin U‐8 a long‐lasting anorectic activity.Subcutaneous injection of PEGylated neuromedin U‐8 exerted an anti‐obesity effect. ABSTRACT Neuromedin U (NMU) is a neuropeptide found in the brain and gastrointestinal tract. The NMU system has been shown to regulate energy homeostasis by both a central and a peripheral mechanism. Peripheral administration of human NMU‐25 was recently shown to inhibit food intake in mice. We examined the possibility that other NMU‐related peptides exert an anorectic activity by intraperitoneal (i.p.) administration. We found that rat NMU‐23 and its structurally‐related peptide rat neuromedin S (NMS) significantly reduced food intake in lean mice, whereas NMU‐8, an active fragment of the octapeptide sequence conserved in porcine, human and mouse NMU, had no effect. When rat NMU‐23, NMU‐8, and rat NMS were covalently conjugated to polyethylene glycol (PEG) (PEGylation) at the N‐terminus of these peptides, PEGylated NMU‐8 showed the most long‐lasting and robust anorectic activity. The exploration of the linker between NMU‐8 and PEG using hetero‐bifunctional chemical cross‐linkers led to an identification of PEGylated NMU‐8 analogs with higher affinity for NMU receptors and with more potent anorectic activity in lean mice. The PEGylated NMU‐8 showed potent and robust anorectic activity and anti‐obesity effect in diet‐induced obesity (DIO) mice by once‐daily subcutaneous (s.c.) administration. These results suggest that PEGylated NMU‐8 has the therapeutic potential for treatment of obesity.

Differential effects of selective agonists of neuromedin U1 and U2 receptors in obese and diabetic mice

Neuromedin U (NmU) may be a novel target for obesity treatment owing to its anorectic and energy expenditure enhancing effects. Although two receptors, NMU1 and NMU2, are both responsible for the NmU‐mediated anti‐obesity effects, the receptor agonist with the most appropriate profiles for treating obesity and diabetes in terms of efficacy and safety is as yet unknown. Thus, we developed and evaluated novel NMU1/2 receptor‐selective agonists.