, % i!¸¸ I N V E S T I G A T I VE U I I O L O ( ; ¥
C()N I E N T A N D DISTR
BU
OF \,ASOA.JTtVE
INTESTINAL POIXPEPTIDE (VIP) !N CAVEBNOUS
L SUL O F H U M A N P E N I S '
MASAFUMI SHII{AI, M.D.
NOBORU ~7\NAIt{ARA, PH.D,
AKtO MAKI, M.D. MASAHAFtU TAKANAMI~ M.D.
CHIZUKO "i%NAIHAt~A, PHD~ KAZUAKI [GUCHI, P~i.D.
KO ANDO, M.D. K I M I I C H I NAKAMURA, M.D.
TSUNEO FUJTIA, M,D. TOSHIHIKO IWANAGA, M.D.
From the Department of Urology and 1st Departmen~ of Internal Medicine, Ibho University School of Medicine. %kyo; the Department of Bioorganic Chemistry, Shizuoka College of Pharmae> Shizuoka; and the Department o:f Anatomy, Niigata University School of Medicine, Niigata, Japan
ABSTBA CT~-J:~eniIe erection is co,ltrolted by a ~atvular structure in the helicine artery in The opening and closing of this valve are believed to be regulated by the autonomic nervou especially through the release of vasoactive i~t('~stinal polTjpeptide (VIY). ~3#~determined th of V1P in cavernous tissue i~ 18 impotent patients and in 5 normat eon~rots b~! radioimm~ and we examined the distributio,z of VIP-ergic nerve fibers i~t ca,)er,wus tissue b?] an imm., chemical method. As a result, it was ;brand that the lower penile VIP content was more among patients with organic impotence than among the controls. Furthermore, VIP-erl fibers u'ere see~, to be di[fuseh,! arid loosely distributed in a large number of orga,~ie h patients. These findings suggest that organic impotence in some patiel#,s' may be d,~e to de~ the VIP content and in VIP-e~gic nerve fibers.
Penile erection is believed to be controlled by a valwflar structure in the helicine artery that adjusls the blood flow into the cavernous spaces of the penis. This vaIvular structure is comprised of smooth muscle tissue and is located at the poi~t: immediately before the helicine arteries open into the cavernous spaces, Further, it has beei~ suggested that the operation of this valve is controlled by the autonomic nervous system, and recently it: has been proposed that this controlling role may be played by the vasoaetive in{estinal polypcptide (VIP)-ergic nerve fibers of the penis. ~ '~ S~ pp( rted in p a n b?, a• 1* ~ r(3 ,lect t:lesearch F t m d from 'Ibho I ) ~ i "*'(rsi~.y,
360
We determined, in normal con impotenk patients, the VIP eonten ous tissue the distribution of VI_P fibers in the cavernous tissue, an level of VIP in the eaverrmus body in both Ji:taceid and erect states. Th~:~ a report of our results. Material and Methods NormaI fresh caverrzous tissu< were obtained from 5 persons, soI were operated on for penis fracture whom had died due to accident or ages of these 5 controls averaged with an age spread o}: sixteet~ ~c
itlOI,OGY
AP[{IL 1990
"
VOI,UM]',
pg/m~ we: weight
........ ssue specimens were also obits w h o u n d e r w e n t silicone iion in the penis as a treatre. Of the 18 impoten~ pacone implants, 3 had been reetion test as having funeFhese 3 patients strongly rentat:ion operation; because ~sponse to various forms of td because these patients, ty-three, and lift:y-nine, saw e for i m p r o v e m e n t of their teed to perform the opera~g 15 patients (average age t age spread of 27 to 67) h a d , due to diabetes mellitus, !raeture with a peIvic bone for cancer of the rectum, or r bladder cancer. Cavernous :~re obtained from the root of ~. One part: of the cavernous s used to measure VIE conn d e r was used to determine VIP-ergic nerve fibers. T h e :~re frozen immediately after tow - 20 °C until the time of nens used in the determinalerve fiber distribution were formalin with a 0.1M phos7,4) for about b,vetve hours. y were immersed in a 30 % vernight, The tissue blocks in liquid nitrogen and cut ice'; in a eryostat, The cryoh e n proeessed by the perox;e (D\P) method ~ using anti] to 1:2,000. The anti-ViP [ was raised against synthetic was shown to reeogifize the VIE v For checking the speeianoreaetion, antiserurn was antigen (I0 ~G/rnt, diluted tory-four hours at 4°(7. The did not show any i m m u n o ;dons examined, These stain~re performed concurrently e conditions, for comparison ment of the content of VIP in ~hed tissue was homogenized of 0.1 M acetic', acid in a horn) at 2°C and then boiled ,?he homogenate so obtained e bath and glacial acetic acid
AP}/IL 1990
/
VOLUME XXXV, NUMBEB 4
2
5O "
t
[
[i
t00
50
C o!"it r'o# t;=:5
cr'~a~ ic impotence n =1 5
fu:acl [:)na'~ impotence r>=:=:3
Fict.:~,'£ !. VtP cor~tent q/caverno~s tis'sue of aH three grou.p~'. was added until aeetic acid contei~ reached 1M. After ten minutes of thorough stirring, the mixture was centri[uged at 28,000 C at 4°C for thirty minutes, and the supernatant was separated, After two volumes o:f IM acetic acid was added to the residue and stirred, centrifugation was done ix~ the same manner, and the resultant superrmtant: was combined with the previous supernatant and used as the e×tracts for examinations. The extracts were [yophi!ized and kept at 4 ~'C until anaIysis. Blood was drawn from the cavernous body in 7 patients with funetional impo~en,ee in a flaccid penile state and from 2 patients (aged 58 and 74) vvi~o became erect at the start: of epidural anesthesia for transurethral resection of d~e prostate as a treatment for prostatic hypertroph> In these patients blood was d r a w n u.i~:}l a syringe containing trace amotmes of heparin and trasyloI and centrifuged i m m e d i a t d > The sela, so separated, were preserved below - 20 °C until used. The VIP content in cavernous tissue or serum was deiermined by radioimmunoassav, s . usino',~synthetic, anti-VIP serum (R
s02). Results VIP contempt i~ cavernous tissue (Fig. l) The VIP content in caverno~ls dss{le in the 5 co~rols ranged from 3!.£ to 81.9 p g / m g wet
361
weight, and the mean content was 60.6 :~:: 19.2 pg/mg wet weight. The VIP content in eaverr> ous tissue in the 3 patienls with fimetional im~ potence was 151.5 pg/mg, 146.1 p g / m g , and I30.0 pg/mg, for a mean value of 142,5 ± 11.2 pg/mg, which was significantly higher than d-mr of the controls (P < 0.001). On the other hand, the corresponding values for the t5 patients with organic impotence ranged from 2.2 p g / m g to 95.0 pg/mg. The VIP content in this group w~us below 10 p g / m g in 5 patients and below 30 p g / m g in 4 patients. In spite of such low values in 9 pat:tents, the mean value for the entire group was 35.3 ± 31.0 p g / m g , which was not: significantly different from that: of the control group. Distributim~ q~ I/1F-e,gic ner,~e _fibers in ca~:ernous tissue
N u m e r o u s n e r v e fibers in t h e c o r p o r a cavernosa of normal subjects stained positively using the anti-VIP serum. VIP-immnnoreactive nerve fibers had a beaded appearance and were densely distributed both in the trabeeulae and in the adventitia of the arteries. A denser innervation of VIP-immunopositive nerves was seen in arterioles having a diameter of 50--150 ~m; d~ese arterioles often followed a helical course (Fig. 2A). In some impotent subjects, VIP-imm u n o r e a c t i v e nerve fibers in t h e c o r p o r a eavernosa remained nmnerous in the trabecutae, while ~hey were few around the arterioles (Fig. 2B). In the other impotent cases, VIP nerve fibers almost completely disappeared throughout the cavernous tissues (Fig. 2C).
'% ,£tt.
~ i
._~
..x.
~
5d
S
t
%,,,
,B
A'
V t P in b l o o d of c a v e r n o u s bod~! o f penis
The leveI of VIP in the blood of flaccid-state penes were undeteetably low in all 7 patients, while 2 patients in an erect state showed elevation in the levels of VIP in ~he blood. The quantities in the latter cases were 8.1 p g / m L (58year-old patient) and 35.0 p g / m I , (74-year-old patient). ( , O r/1 ttl (YI
As we m e n t i o n e d before, n u m e r o u s VIPergic nerve fibers were seen around the valwflar structure of the helicine artery in the h u m a n penile cavernous tissue. This fact suggests that VIP-ergic nerve fibers are invoh~ed in the opening and closing of d~e helieine artery valve. On the other hand, the content of VIP in penile cavernous tissue was quite high, while VIP in the blood from penile cavernous tissue in a flac-
~2
C
;;-
F~c;t:~: 2. 1)isH'ibutim~ of Vlf~-et~g#: nerc, e /fbes ca~er'nou.s li.s~sue, t'At-s~ammg r.~>iH~use ~)f an~i.serum. (A) Normal eor~roh VIP-i,,)~mm~oreaG ner~e fibers h,ave beaded ,s~r**ct,~re, a,r~ dea, distributed around tu',o a~'~erioles (A % and fol helical co,~r.s'es. S: caver,~ou, si,ms'. ('B) a~.d Diabetic i'mpote~,ce: (B) VIt~.-fmm~.e~.oreacli ~e r~ Jiber,~ are stilI numerous 'i~ frabecMae; (C) fibers ,wiflt au!! dis:tinct "" "~ could be s'~c~t i~ eilher frabec.utac or arou~.~c, arteriole (A'). S; caver~o~, .vhms. "
eid state was undeteetabte. Int.e the state of the penis changes t erect, the blood level of VIP in tissue becomes elevated. This c led us to postulate that VIE wi
UROIZ)(;Y
APRIL 1990
VOL*, MI? XXX\'I
~rve fibers just before erection, muscle tissue of the h d i e i n e ax, thereby opening the valve. ed VIP in the penile cavernous to Ottesen and co-workers 9 an nto the penile cavernous tissue erection. It therefore seems ' plays an i m p o r t a n t role in ld distribution of VIP in caw not differ between the impothe controls, while the content ous tissue was elevated in pa~mal impotence. This fact may by assuming that. VIP is for efficiently utilized in rune: patients. As ours was only a e cause of this remains to be lrther studies involving m u c h f patiems. nt in cavernous tissue was low w i t h organic impotence, and imens stained poorly in im:al studies. fimens were stained concurthe same conditions, but since .s histologie, not quantitative,
APllIL 1990
/
VOLEME XXXV; NUMBEB 4
we cannot make any quar~fitative eomparisons; howevei, it is interesting to note ~:hat the results of stai~fing were etosely c o r r d a t e d with the VIP content in eaverrlous tissue. We plan further studies to substantiate this relationship between staining and the VIP content of cavernous t:isst~e of the penis. Tokyo 1.43, Japan (DR. SmaAI) References 1. Polak .~M, Cu J, Mina S, ai~d Bloom SR: VIPergic aerv~-s in the pe~is, Lancet 2:217 (t9,$I). 2, Virag 1"~, et aI: \~asoae~ive i~testi~la! po~ypeptide rdeas~,: during penile erection in man, Lancet 2:11{~8 (1q82). 3. Gu J, etaI: Peptidergic L'mervatioa of the }roman genital traeL J Urol 130:386 (1983). 4. Crowe B., et aI: ~';:isoac,tive in{estictal polypeptidedike immunoreactive nerves in diabetic penis. A comparison between streptozntoein-treated raks and man, Diabetes 32:1075 (1983), 5. Cu J, et al: Decrease of vasoadive intestiual polypeptide (VIP) in the penis~ from impotent men, Lancet 2:315 (I984). 6, Sternberger LA: [mmunob.istochemistry, 2~.d ed., New Y~rk, Joh~ Wiley and Sons, 1979, p i 7. ?~maihara C~ et al: hm~mnoreadive VIP (vasoactive int{:
C()N I E N T A N D DISTR
BU
OF \,ASOA.JTtVE
INTESTINAL POIXPEPTIDE (VIP) !N CAVEBNOUS
L SUL O F H U M A N P E N I S '
MASAFUMI SHII{AI, M.D.
NOBORU ~7\NAIt{ARA, PH.D,
AKtO MAKI, M.D. MASAHAFtU TAKANAMI~ M.D.
CHIZUKO "i%NAIHAt~A, PHD~ KAZUAKI [GUCHI, P~i.D.
KO ANDO, M.D. K I M I I C H I NAKAMURA, M.D.
TSUNEO FUJTIA, M,D. TOSHIHIKO IWANAGA, M.D.
From the Department of Urology and 1st Departmen~ of Internal Medicine, Ibho University School of Medicine. %kyo; the Department of Bioorganic Chemistry, Shizuoka College of Pharmae> Shizuoka; and the Department o:f Anatomy, Niigata University School of Medicine, Niigata, Japan
ABSTBA CT~-J:~eniIe erection is co,ltrolted by a ~atvular structure in the helicine artery in The opening and closing of this valve are believed to be regulated by the autonomic nervou especially through the release of vasoactive i~t('~stinal polTjpeptide (VIY). ~3#~determined th of V1P in cavernous tissue i~ 18 impotent patients and in 5 normat eon~rots b~! radioimm~ and we examined the distributio,z of VIP-ergic nerve fibers i~t ca,)er,wus tissue b?] an imm., chemical method. As a result, it was ;brand that the lower penile VIP content was more among patients with organic impotence than among the controls. Furthermore, VIP-erl fibers u'ere see~, to be di[fuseh,! arid loosely distributed in a large number of orga,~ie h patients. These findings suggest that organic impotence in some patiel#,s' may be d,~e to de~ the VIP content and in VIP-e~gic nerve fibers.
Penile erection is believed to be controlled by a valwflar structure in the helicine artery that adjusls the blood flow into the cavernous spaces of the penis. This vaIvular structure is comprised of smooth muscle tissue and is located at the poi~t: immediately before the helicine arteries open into the cavernous spaces, Further, it has beei~ suggested that the operation of this valve is controlled by the autonomic nervous system, and recently it: has been proposed that this controlling role may be played by the vasoaetive in{estinal polypcptide (VIP)-ergic nerve fibers of the penis. ~ '~ S~ pp( rted in p a n b?, a• 1* ~ r(3 ,lect t:lesearch F t m d from 'Ibho I ) ~ i "*'(rsi~.y,
360
We determined, in normal con impotenk patients, the VIP eonten ous tissue the distribution of VI_P fibers in the cavernous tissue, an level of VIP in the eaverrmus body in both Ji:taceid and erect states. Th~:~ a report of our results. Material and Methods NormaI fresh caverrzous tissu< were obtained from 5 persons, soI were operated on for penis fracture whom had died due to accident or ages of these 5 controls averaged with an age spread o}: sixteet~ ~c
itlOI,OGY
AP[{IL 1990
"
VOI,UM]',
pg/m~ we: weight
........ ssue specimens were also obits w h o u n d e r w e n t silicone iion in the penis as a treatre. Of the 18 impoten~ pacone implants, 3 had been reetion test as having funeFhese 3 patients strongly rentat:ion operation; because ~sponse to various forms of td because these patients, ty-three, and lift:y-nine, saw e for i m p r o v e m e n t of their teed to perform the opera~g 15 patients (average age t age spread of 27 to 67) h a d , due to diabetes mellitus, !raeture with a peIvic bone for cancer of the rectum, or r bladder cancer. Cavernous :~re obtained from the root of ~. One part: of the cavernous s used to measure VIE conn d e r was used to determine VIP-ergic nerve fibers. T h e :~re frozen immediately after tow - 20 °C until the time of nens used in the determinalerve fiber distribution were formalin with a 0.1M phos7,4) for about b,vetve hours. y were immersed in a 30 % vernight, The tissue blocks in liquid nitrogen and cut ice'; in a eryostat, The cryoh e n proeessed by the perox;e (D\P) method ~ using anti] to 1:2,000. The anti-ViP [ was raised against synthetic was shown to reeogifize the VIE v For checking the speeianoreaetion, antiserurn was antigen (I0 ~G/rnt, diluted tory-four hours at 4°(7. The did not show any i m m u n o ;dons examined, These stain~re performed concurrently e conditions, for comparison ment of the content of VIP in ~hed tissue was homogenized of 0.1 M acetic', acid in a horn) at 2°C and then boiled ,?he homogenate so obtained e bath and glacial acetic acid
AP}/IL 1990
/
VOLUME XXXV, NUMBEB 4
2
5O "
t
[
[i
t00
50
C o!"it r'o# t;=:5
cr'~a~ ic impotence n =1 5
fu:acl [:)na'~ impotence r>=:=:3
Fict.:~,'£ !. VtP cor~tent q/caverno~s tis'sue of aH three grou.p~'. was added until aeetic acid contei~ reached 1M. After ten minutes of thorough stirring, the mixture was centri[uged at 28,000 C at 4°C for thirty minutes, and the supernatant was separated, After two volumes o:f IM acetic acid was added to the residue and stirred, centrifugation was done ix~ the same manner, and the resultant superrmtant: was combined with the previous supernatant and used as the e×tracts for examinations. The extracts were [yophi!ized and kept at 4 ~'C until anaIysis. Blood was drawn from the cavernous body in 7 patients with funetional impo~en,ee in a flaccid penile state and from 2 patients (aged 58 and 74) vvi~o became erect at the start: of epidural anesthesia for transurethral resection of d~e prostate as a treatment for prostatic hypertroph> In these patients blood was d r a w n u.i~:}l a syringe containing trace amotmes of heparin and trasyloI and centrifuged i m m e d i a t d > The sela, so separated, were preserved below - 20 °C until used. The VIP content in cavernous tissue or serum was deiermined by radioimmunoassav, s . usino',~synthetic, anti-VIP serum (R
s02). Results VIP contempt i~ cavernous tissue (Fig. l) The VIP content in caverno~ls dss{le in the 5 co~rols ranged from 3!.£ to 81.9 p g / m g wet
361
weight, and the mean content was 60.6 :~:: 19.2 pg/mg wet weight. The VIP content in eaverr> ous tissue in the 3 patienls with fimetional im~ potence was 151.5 pg/mg, 146.1 p g / m g , and I30.0 pg/mg, for a mean value of 142,5 ± 11.2 pg/mg, which was significantly higher than d-mr of the controls (P < 0.001). On the other hand, the corresponding values for the t5 patients with organic impotence ranged from 2.2 p g / m g to 95.0 pg/mg. The VIP content in this group w~us below 10 p g / m g in 5 patients and below 30 p g / m g in 4 patients. In spite of such low values in 9 pat:tents, the mean value for the entire group was 35.3 ± 31.0 p g / m g , which was not: significantly different from that: of the control group. Distributim~ q~ I/1F-e,gic ner,~e _fibers in ca~:ernous tissue
N u m e r o u s n e r v e fibers in t h e c o r p o r a cavernosa of normal subjects stained positively using the anti-VIP serum. VIP-immnnoreactive nerve fibers had a beaded appearance and were densely distributed both in the trabeeulae and in the adventitia of the arteries. A denser innervation of VIP-immunopositive nerves was seen in arterioles having a diameter of 50--150 ~m; d~ese arterioles often followed a helical course (Fig. 2A). In some impotent subjects, VIP-imm u n o r e a c t i v e nerve fibers in t h e c o r p o r a eavernosa remained nmnerous in the trabecutae, while ~hey were few around the arterioles (Fig. 2B). In the other impotent cases, VIP nerve fibers almost completely disappeared throughout the cavernous tissues (Fig. 2C).
'% ,£tt.
~ i
._~
..x.
~
5d
S
t
%,,,
,B
A'
V t P in b l o o d of c a v e r n o u s bod~! o f penis
The leveI of VIP in the blood of flaccid-state penes were undeteetably low in all 7 patients, while 2 patients in an erect state showed elevation in the levels of VIP in ~he blood. The quantities in the latter cases were 8.1 p g / m L (58year-old patient) and 35.0 p g / m I , (74-year-old patient). ( , O r/1 ttl (YI
As we m e n t i o n e d before, n u m e r o u s VIPergic nerve fibers were seen around the valwflar structure of the helicine artery in the h u m a n penile cavernous tissue. This fact suggests that VIP-ergic nerve fibers are invoh~ed in the opening and closing of d~e helieine artery valve. On the other hand, the content of VIP in penile cavernous tissue was quite high, while VIP in the blood from penile cavernous tissue in a flac-
~2
C
;;-
F~c;t:~: 2. 1)isH'ibutim~ of Vlf~-et~g#: nerc, e /fbes ca~er'nou.s li.s~sue, t'At-s~ammg r.~>iH~use ~)f an~i.serum. (A) Normal eor~roh VIP-i,,)~mm~oreaG ner~e fibers h,ave beaded ,s~r**ct,~re, a,r~ dea, distributed around tu',o a~'~erioles (A % and fol helical co,~r.s'es. S: caver,~ou, si,ms'. ('B) a~.d Diabetic i'mpote~,ce: (B) VIt~.-fmm~.e~.oreacli ~e r~ Jiber,~ are stilI numerous 'i~ frabecMae; (C) fibers ,wiflt au!! dis:tinct "" "~ could be s'~c~t i~ eilher frabec.utac or arou~.~c, arteriole (A'). S; caver~o~, .vhms. "
eid state was undeteetabte. Int.e the state of the penis changes t erect, the blood level of VIP in tissue becomes elevated. This c led us to postulate that VIE wi
UROIZ)(;Y
APRIL 1990
VOL*, MI? XXX\'I
~rve fibers just before erection, muscle tissue of the h d i e i n e ax, thereby opening the valve. ed VIP in the penile cavernous to Ottesen and co-workers 9 an nto the penile cavernous tissue erection. It therefore seems ' plays an i m p o r t a n t role in ld distribution of VIP in caw not differ between the impothe controls, while the content ous tissue was elevated in pa~mal impotence. This fact may by assuming that. VIP is for efficiently utilized in rune: patients. As ours was only a e cause of this remains to be lrther studies involving m u c h f patiems. nt in cavernous tissue was low w i t h organic impotence, and imens stained poorly in im:al studies. fimens were stained concurthe same conditions, but since .s histologie, not quantitative,
APllIL 1990
/
VOLEME XXXV; NUMBEB 4
we cannot make any quar~fitative eomparisons; howevei, it is interesting to note ~:hat the results of stai~fing were etosely c o r r d a t e d with the VIP content in eaverrlous tissue. We plan further studies to substantiate this relationship between staining and the VIP content of cavernous t:isst~e of the penis. Tokyo 1.43, Japan (DR. SmaAI) References 1. Polak .~M, Cu J, Mina S, ai~d Bloom SR: VIPergic aerv~-s in the pe~is, Lancet 2:217 (t9,$I). 2, Virag 1"~, et aI: \~asoae~ive i~testi~la! po~ypeptide rdeas~,: during penile erection in man, Lancet 2:11{~8 (1q82). 3. Gu J, etaI: Peptidergic L'mervatioa of the }roman genital traeL J Urol 130:386 (1983). 4. Crowe B., et aI: ~';:isoac,tive in{estictal polypeptidedike immunoreactive nerves in diabetic penis. A comparison between streptozntoein-treated raks and man, Diabetes 32:1075 (1983), 5. Cu J, et al: Decrease of vasoadive intestiual polypeptide (VIP) in the penis~ from impotent men, Lancet 2:315 (I984). 6, Sternberger LA: [mmunob.istochemistry, 2~.d ed., New Y~rk, Joh~ Wiley and Sons, 1979, p i 7. ?~maihara C~ et al: hm~mnoreadive VIP (vasoactive int{:
