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511 Role of central tachykinin peptides in water balance of rats
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509
IMMUNOHISTOCHEMICAL S T U D Y OF P O S T N A T A L DEVELOPMENT DELTORPHIN-I IN THE RAT BRAIN. KAEU0 NONOMURA 1~
OF IKUO
[ D - A I a 2] TOOYA~A2~
HIROMICHI~BE 2 ,_TINDARO_RENDA3j__3ZITTQRIO E R S P A M E R 4 ~ ~ I R Q S H I KIMURA 2~ IDept. of Pediatric .__anct_21nst._of_Moi._Neurobiol.,~higa_Mniv~_Df M~d~__Sc/~ Qts/1, 520-21 Japan;3Inst._of_Human_Anat.~and_4Emeritus of Ned. Pharmacol., un/xers/ty "La Sapienza2" of_Rome ,_Italy Postnatal dcvclopmcnt of [D-Ala2]-deltorphin 1 (DADTI), a highly selective ligand for 6 opioid receptors, was studied in the rat brain by immunohistochemistry using a specific antiserum. DADTI-positive staining occurred mainly in neurons, and occasionally in tanycytes but never in glia cclls. On postnatal day 0 (P0), positive neuronal somata were distributed in specific brain regions including the olfactory tubercle, ventral pallidum, hippocampus, ventral tegmental area, pars compacta of the substantia nigra, supramammillary nucleus, and dorsal raphe nucleus. Positive nerve fibers were obsen'ed in the external plexiform layer of the main olfactory bulb, accessory olfactory bulb, olfactory tubercle, prefrontal cortex, cingulate cortex, neostriatum, accumbens, lateral scptal nucleus, lateral habenular nucleus, and superior colliculus. Positive cell bodies were decreased gradually with age in both density and staining intensity, and those in the olfactory tubercle and ventral pallidum were no longer visible by P7 and PI4, respectively. Positive fibers were also decreased in density with age except" for the olfactory tubercle. The results suggest that DADTI-Iikc molecule(s) play an important role on the development of certain neuronal systems.
510
DEVELOPMENT OF CHEMILUMINESCENCE ASSAY FOR PREPROCHOLECYSTOKININ MRNA AND ITS APPLICATION SHINJI TAKAHASHI1, TAKAYUKI YAMASHITA l, KATUMICHI TAKEDA2, MICHIE OTO 3 AND TAKESHI KATO l, ILa_b.of Molecular Recognition, Graduate Sch. of Integrated Sci., Yokohama City Univ., 2Research and Development Center, DAINABOT CO., LTD, 3Dep. of Biotech., Tokyo Technical College Developmental change of preprocholecystokinin mRNA (CCK mRNA) was examined in rat brain regions using chemiluminescence method. Northern blot analysis of total RNA from the rat brain regions at 6 weeks of age showed that there were no other bands except CCK mRNA and the CCK mRNA level in the frontal cortex was higher than the level in the hippocampus. Dot blot analysis could detect 0.312 p.g of total RNA from the frontal cortex at 6 weeks of age. In the hippocampus at 6 weeks of age, the CCK mRNA level was approximately 40 % of the level in the frontal cortex. At a day of age, the CCK mRNA level in the frontal cortex was 20-25 % of the 6 weeks of age level. These results are consistent with our previous study using RI, suggesting that this chemiluminescence method is appropriate to detection of mRNA without using RI. We are going to apply this method to further study about regulation mechanism of CCK mRNA expression.
511
ROLE OF CENTRAL TACHYKININ PEPTIDES 1N WATER BALANCE OF RATS. AKI SAIGO1, YUKIO TAKANO t, TAKASI EGUCHI],TAKAFUMI MATSUMOTO 1, YASUHISA NAKAYAMA 1, RYO SAITO 1, KASTUSHI YAMADA2 AND HIRO-O KAMIYA1, tDepartment of Pharmacology, Faculty of Pharmaceutical Sciences, 2School of medisine, Fukuoka University, Fukuoka 814-01, Japan.
In the present study, we examined the effects of central tachykinin peptides on water intake and an antidiuretic action in conscious rats. Intracerebreventricular (i.c.v.) injections of tachykinin NK-1 agonist, substance P (SP) (0.1 - 1 nmol), and the selective NK-3 agonist, senktide (0.01 - 1 nmol) inhibited water intake induced by angiotensin II (100 ng, i.c.v.), isoproterenol (200 mg/kg, s.c.), hypertonic NaCI (s.c.) and 48 hr water deprivation respectively. Inhibitory effects of senktide on experimentally-stimulated water intake was much more potent than SP. Senktide (0.01 - 10 nmol, i.c.v.) resulted in prolonged inhibition of urine output, but SP did not inhibit urine output in water-loaded rats (4.5 % body weight). The antidiuretic effect of senktide was inhibited by pretreatment with a non-peptide vasopressin V2 antagonist, OPC-31260 (1 mg/kg, i.v.). In addition, senktide (0.1 - 1 nmol, i.c.v.) caused a marked increase of the plasma AVP level in conscious waterloaded rats. These results suggest that the central.tachykinin peptides, especially the NK-3 agonists, play a role in the central regulation of water balance. Ref:Neurosci. Lett. in press (1993). Reg. Peptides 46 (1993) 360.
509
IMMUNOHISTOCHEMICAL S T U D Y OF P O S T N A T A L DEVELOPMENT DELTORPHIN-I IN THE RAT BRAIN. KAEU0 NONOMURA 1~
OF IKUO
[ D - A I a 2] TOOYA~A2~
HIROMICHI~BE 2 ,_TINDARO_RENDA3j__3ZITTQRIO E R S P A M E R 4 ~ ~ I R Q S H I KIMURA 2~ IDept. of Pediatric .__anct_21nst._of_Moi._Neurobiol.,~higa_Mniv~_Df M~d~__Sc/~ Qts/1, 520-21 Japan;3Inst._of_Human_Anat.~and_4Emeritus of Ned. Pharmacol., un/xers/ty "La Sapienza2" of_Rome ,_Italy Postnatal dcvclopmcnt of [D-Ala2]-deltorphin 1 (DADTI), a highly selective ligand for 6 opioid receptors, was studied in the rat brain by immunohistochemistry using a specific antiserum. DADTI-positive staining occurred mainly in neurons, and occasionally in tanycytes but never in glia cclls. On postnatal day 0 (P0), positive neuronal somata were distributed in specific brain regions including the olfactory tubercle, ventral pallidum, hippocampus, ventral tegmental area, pars compacta of the substantia nigra, supramammillary nucleus, and dorsal raphe nucleus. Positive nerve fibers were obsen'ed in the external plexiform layer of the main olfactory bulb, accessory olfactory bulb, olfactory tubercle, prefrontal cortex, cingulate cortex, neostriatum, accumbens, lateral scptal nucleus, lateral habenular nucleus, and superior colliculus. Positive cell bodies were decreased gradually with age in both density and staining intensity, and those in the olfactory tubercle and ventral pallidum were no longer visible by P7 and PI4, respectively. Positive fibers were also decreased in density with age except" for the olfactory tubercle. The results suggest that DADTI-Iikc molecule(s) play an important role on the development of certain neuronal systems.
510
DEVELOPMENT OF CHEMILUMINESCENCE ASSAY FOR PREPROCHOLECYSTOKININ MRNA AND ITS APPLICATION SHINJI TAKAHASHI1, TAKAYUKI YAMASHITA l, KATUMICHI TAKEDA2, MICHIE OTO 3 AND TAKESHI KATO l, ILa_b.of Molecular Recognition, Graduate Sch. of Integrated Sci., Yokohama City Univ., 2Research and Development Center, DAINABOT CO., LTD, 3Dep. of Biotech., Tokyo Technical College Developmental change of preprocholecystokinin mRNA (CCK mRNA) was examined in rat brain regions using chemiluminescence method. Northern blot analysis of total RNA from the rat brain regions at 6 weeks of age showed that there were no other bands except CCK mRNA and the CCK mRNA level in the frontal cortex was higher than the level in the hippocampus. Dot blot analysis could detect 0.312 p.g of total RNA from the frontal cortex at 6 weeks of age. In the hippocampus at 6 weeks of age, the CCK mRNA level was approximately 40 % of the level in the frontal cortex. At a day of age, the CCK mRNA level in the frontal cortex was 20-25 % of the 6 weeks of age level. These results are consistent with our previous study using RI, suggesting that this chemiluminescence method is appropriate to detection of mRNA without using RI. We are going to apply this method to further study about regulation mechanism of CCK mRNA expression.
511
ROLE OF CENTRAL TACHYKININ PEPTIDES 1N WATER BALANCE OF RATS. AKI SAIGO1, YUKIO TAKANO t, TAKASI EGUCHI],TAKAFUMI MATSUMOTO 1, YASUHISA NAKAYAMA 1, RYO SAITO 1, KASTUSHI YAMADA2 AND HIRO-O KAMIYA1, tDepartment of Pharmacology, Faculty of Pharmaceutical Sciences, 2School of medisine, Fukuoka University, Fukuoka 814-01, Japan.
In the present study, we examined the effects of central tachykinin peptides on water intake and an antidiuretic action in conscious rats. Intracerebreventricular (i.c.v.) injections of tachykinin NK-1 agonist, substance P (SP) (0.1 - 1 nmol), and the selective NK-3 agonist, senktide (0.01 - 1 nmol) inhibited water intake induced by angiotensin II (100 ng, i.c.v.), isoproterenol (200 mg/kg, s.c.), hypertonic NaCI (s.c.) and 48 hr water deprivation respectively. Inhibitory effects of senktide on experimentally-stimulated water intake was much more potent than SP. Senktide (0.01 - 10 nmol, i.c.v.) resulted in prolonged inhibition of urine output, but SP did not inhibit urine output in water-loaded rats (4.5 % body weight). The antidiuretic effect of senktide was inhibited by pretreatment with a non-peptide vasopressin V2 antagonist, OPC-31260 (1 mg/kg, i.v.). In addition, senktide (0.1 - 1 nmol, i.c.v.) caused a marked increase of the plasma AVP level in conscious waterloaded rats. These results suggest that the central.tachykinin peptides, especially the NK-3 agonists, play a role in the central regulation of water balance. Ref:Neurosci. Lett. in press (1993). Reg. Peptides 46 (1993) 360.