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Tracheal dilation induced by posterior hypothalamic stimulation
SlOX
3-40
IMMUNOHISTOCHEMICAL
DEMONSTRATION
OF POLYAMINES
IN THE BRAIN. REIJI SEMBA,
Department of Anatomy, Mie University School of Medicine, Tsu, Mie 514, Japan Polyamines are believed to play important roles in controlling the cellular growth. A considerable
amount of
polyamines are present in the brain and a new role of polyamines in the brain as a modulator of NMDA receptor is also suggested.
However, there is no information so far on the precise localization of polyamines.
against polyamines and use it for demonstrating immunohistochemical chromatographies. anti-spermine
method. We raised anti-polyamine Anti-putrescine
antisera in rabbits and purified antibodies using affinity
antibody was crossreactive with spermidine but not with spermine, and anti-spermidine
antibodies were crossreactive
immunohistochemical
We attempted to raise antibodies
the localization of polyamines on histological sections by an
to each other but not with putrescine.
localization of polyamines in the brain as well as in other tissues.
polyamines were found to be concentrated
and
These antibodies enabled us to study the
in astrocytes and in some particular neurons.
In the central nervous system. Present results suggest new roles of
polyamines in the glial cells and these particular neurons.
3-41
TKACf1EAL DILATION INDlICED BY f'OSTERJOR IIYPOI'HAf.AMI(:STl?lUI,ATION. \lICHIKO -1 IiGASE' MITSUKO HIRUKAWA' IKUO HOMMAI, SEIJL SHIODA' AND YhSU4lITSU KAK.41;::'. DepartmentP of 'Physiology & 'Anatomy, _SGowaiversity SchoolofMedicine, Shinagawa-ku, Tokyo 142, Japan. -We have previously reported histamine immunoreactive (HA-I) cell bodies distributed in the tuberomammillary nucleus (TMK;) of the posterior hypothalamus and fibers in the medulla ohIungat,l, in rabbit. Application of HA to the nucleus of the sol it.ary trt3c.t(NTS) caused t r-acheal di Idt ii,!!. 'The present study was to examine t.racheal pressure change induced t,> posterior hypot!-ialamr Fine structure and synaptic regulatiun of HA-Ir trachea. Tracheal d i tat ion was i ntiuccd frequency dependently by electrical stimulation (0.5 msec pulses, i-100 pps, 75-200 uA) 01 the, Application of pyrilamine (250 nmol) into the fourth ventricle reduced this dilation TO-50%. TMK. For immunohistochemical study, the rabbits were anesthetized witI pentobarbit~al, and per t L,S~‘d transcardial 1 y with 4% I-ethyl-3-(H-dimethyl-aminof)rnppl)carbodi imide (w/v), fol lowed b) ,‘“/, glutaraldehyde (v/v). Sections cuL in a vibratome were’ immunosta ined by the ABC method us i 11s HA antibody. HA-1 axon terminals were observed to synapse in the NTS. Results suggest that ffA-canta i n i ng neurons that or ig i nat R iI,tI~<~ '1'MY modulalr Ii-;lrhe,rI [)!-t~s~ur(~ through hi stami “e-H, receptors at the level (,f li~c‘ medr~l I,]ohloi?p;~t,i.
3-42
"ENTRAL
VENTRICLE HOplilA, --_-_ Hatanodai,
EFFECTS
OF
HISTA:1IldE
ON
IN A!qESTHETIZED RASBIT.
Department Shinagawa-ku,
of
Physiology, Tokyo 142,
TRACI-!EAL
YITSUKO
!: APPLIC3 TO T :H E F 0 u ii'I' HIRUKAI'JA ‘lICHIK0 1'14SE AND ':CUO L__________I-L____:__.z___
PRESSURE
School-i-~~~icine--Chowa I_-~______________Y--z-z_ Universit Japan
150
Immuno~~istochemical studies have shown tk$;athistamine (HA) contaln~ns I1c:urons were distributed in the rabbit brain stem. Correlation between the central effects of HA and tracheal pressure (P,) was examined in anesthetized rabbits. 'iabbits were anesthetized With 450 mg/kg urethane plus 45 mg/ky chloralose l.v., paralyzed w1tt. 1.5 mg/kg gallamine triethiodide i.v., and artificially hyperventilated. Rddltlondl urethane and chloralose, or gallamine triethiodide was administered as neede.d. 15 balloon was inserted into the upper trachea to measure PT through a L3res s u re transducer. The fourth ventricle (IV) was exposed dorsally. HA (1.5, 5.-o, 15.0 nmol), pyrilamine (250 nmol) and cimetidine (400 nmol) were dissolved ln artlflcial cerebrospinal fluid and adjusted to pH 7.0 - 7.4. The 10 ~1 compound solutiori was injected into the IV with a micropipette. HA applied to the IV dose-dependently decreased P soon after administration with recovery 2-5 mln later. The tracheal dilation in X'uced by i_TAwas blocked by pyrilarnlne injected Into the IV 15 mln before the HA administration, but was not blocked by clmetidlne. The results show cerltral effects of HA on traclieal dilation through histamine-s, receptors I li t r-1,: anesthetized rabbit.
3-40
IMMUNOHISTOCHEMICAL
DEMONSTRATION
OF POLYAMINES
IN THE BRAIN. REIJI SEMBA,
Department of Anatomy, Mie University School of Medicine, Tsu, Mie 514, Japan Polyamines are believed to play important roles in controlling the cellular growth. A considerable
amount of
polyamines are present in the brain and a new role of polyamines in the brain as a modulator of NMDA receptor is also suggested.
However, there is no information so far on the precise localization of polyamines.
against polyamines and use it for demonstrating immunohistochemical chromatographies. anti-spermine
method. We raised anti-polyamine Anti-putrescine
antisera in rabbits and purified antibodies using affinity
antibody was crossreactive with spermidine but not with spermine, and anti-spermidine
antibodies were crossreactive
immunohistochemical
We attempted to raise antibodies
the localization of polyamines on histological sections by an
to each other but not with putrescine.
localization of polyamines in the brain as well as in other tissues.
polyamines were found to be concentrated
and
These antibodies enabled us to study the
in astrocytes and in some particular neurons.
In the central nervous system. Present results suggest new roles of
polyamines in the glial cells and these particular neurons.
3-41
TKACf1EAL DILATION INDlICED BY f'OSTERJOR IIYPOI'HAf.AMI(:STl?lUI,ATION. \lICHIKO -1 IiGASE' MITSUKO HIRUKAWA' IKUO HOMMAI, SEIJL SHIODA' AND YhSU4lITSU KAK.41;::'. DepartmentP of 'Physiology & 'Anatomy, _SGowaiversity SchoolofMedicine, Shinagawa-ku, Tokyo 142, Japan. -We have previously reported histamine immunoreactive (HA-I) cell bodies distributed in the tuberomammillary nucleus (TMK;) of the posterior hypothalamus and fibers in the medulla ohIungat,l, in rabbit. Application of HA to the nucleus of the sol it.ary trt3c.t(NTS) caused t r-acheal di Idt ii,!!. 'The present study was to examine t.racheal pressure change induced t,> posterior hypot!-ialamr Fine structure and synaptic regulatiun of HA-I
3-42
"ENTRAL
VENTRICLE HOplilA, --_-_ Hatanodai,
EFFECTS
OF
HISTA:1IldE
ON
IN A!qESTHETIZED RASBIT.
Department Shinagawa-ku,
of
Physiology, Tokyo 142,
TRACI-!EAL
YITSUKO
!: APPLIC3 TO T :H E F 0 u ii'I' HIRUKAI'JA ‘lICHIK0 1'14SE AND ':CUO L__________I-L____:__.z___
PRESSURE
School-i-~~~icine--Chowa I_-~______________Y--z-z_ Universit Japan
150
Immuno~~istochemical studies have shown tk$;athistamine (HA) contaln~ns I1c:urons were distributed in the rabbit brain stem. Correlation between the central effects of HA and tracheal pressure (P,) was examined in anesthetized rabbits. 'iabbits were anesthetized With 450 mg/kg urethane plus 45 mg/ky chloralose l.v., paralyzed w1tt. 1.5 mg/kg gallamine triethiodide i.v., and artificially hyperventilated. Rddltlondl urethane and chloralose, or gallamine triethiodide was administered as neede.d. 15 balloon was inserted into the upper trachea to measure PT through a L3res s u re transducer. The fourth ventricle (IV) was exposed dorsally. HA (1.5, 5.-o, 15.0 nmol), pyrilamine (250 nmol) and cimetidine (400 nmol) were dissolved ln artlflcial cerebrospinal fluid and adjusted to pH 7.0 - 7.4. The 10 ~1 compound solutiori was injected into the IV with a micropipette. HA applied to the IV dose-dependently decreased P soon after administration with recovery 2-5 mln later. The tracheal dilation in X'uced by i_TAwas blocked by pyrilarnlne injected Into the IV 15 mln before the HA administration, but was not blocked by clmetidlne. The results show cerltral effects of HA on traclieal dilation through histamine-s, receptors I li t r-1,: anesthetized rabbit.