Effects of haemorrhagic hypotension on the subcapsular artery and microvasculature of the rat testis.

Developing germ cells may be sensitive to even moderate reductions in blood flow. Surprisingly, however, experimental evidence suggests that the rat testis may be unable to maintain its blood flow during a decrease in systemic blood pressure. This study was therefore performed in order to answer the following questions: Is the testis able to maintain its blood flow during moderate to major reductions in blood pressure and, if so, at which level of the testicular vasculature (main artery or microcirculation) does this compensatory response take place? Moderate (-20%) and major (-40%) reductions in blood pressure were induced in anaesthetized rats by haemorrhage and the effects on testicular microvascular blood flow and subcapsular testicular artery diameter were examined by using laser Doppler flowmetry and in vivo video-microscopy respectively. Haemorrhagic hypotension led to decreased local testicular blood flow, but the relative reductions in flow were generally only half as large as the reductions in blood pressure. Hypotension also decreased the diameter of the main subcapsular testicular artery. During large reductions in blood pressure the subcapsular testicular artery constricts and testicular blood flow decreases. However, blood flow is reduced proportionally less than the mean arterial pressure, suggesting that local regulatory mechanisms are present in the testicular microvasculature, which may prevent blood flow from falling below a critical level.

Effects of acute graded reductions in testicular blood flow on testicular morphology in the adult rat.

The effect of moderate reductions in testicular blood flow has not been studied systematically. The aim of this study was, therefore, to examine the effects of different degrees of blood flow reduction on testicular morphology and to determine how much flow can be reduced before damage occurs. The subcapsular testicular artery was partially ligated in the left testes of adult rats. Testicular blood flow was measured before, immediately after, and 5 h after the ligation using laser Doppler flowmetry. After 5 h of partial ligation, the testes were removed, and their morphology was examined and related to the degree of blood flow reduction. The number of in situ end-labeled- or TUNEL-positive (i.e., dying) germ cells and the volume density of intravascular polymorphonuclear (PMN) leukocytes were measured. When flow was reduced to approximately 70% or less of its pretreatment value, a dose-related increase in the number of dying spermatogonia and early spermatocytes was seen. The PMN leukocytes accumulated in testicular blood vessels after partial ligation, and the maximum number was observed in testes where flow was reduced by approximately 50% of the pretreatment value. In conclusion, early stages of spermatogenesis are sensitive to a moderate, acute reduction in blood flow. Discrete reductions in flow may, therefore, have a large impact on sperm production.

Blood vessels are regulators of growth, diagnostic markers and therapeutic targets in prostate cancer.

The vasculature plays an important role in the normal and malignant prostate. Under basal conditions both glandular epithelial and stromal prostate cells produce an abundance of blood flow and angiogenesis regulating substances and the expression of these is generally increased in prostate tumors. The proportion of proliferating endothelial cells is high in the normal prostate compared to other tissues in the body. After castration effects on the vasculature, such as decreased blood flow and vascular regression, precede effects on the glandular compartment. Correspondingly, hormone induced prostate growth is characterized by early effects on the vasculature such as increased blood flow and endothelial cell proliferation, thus indicating that the vasculature may be involved in the androgenic regulation of the prostate. Prostatic intraepithelial neoplasia (PIN) and prostate cancer are associated with increased vascular density and in experimental models prostate cancer growth is apparently angiogenesis-dependent since tumor growth and progression can be inhibited by antiangiogenic treatment. Moreover, vascular density has been related to prognosis in prostate cancer patients. A better understanding of the pathways regulating angiogenesis in the normal prostate and how these pathways change during malignant transformation can hopefully lead to better prognostic markers and therapies for the large group of patients with prostate cancer. The purpose of this review is therefore to summarize the current knowledge on the role and regulation of the vasculature in the prostate and its potential clinical applications.

Temporal variations in testicular microcirculation.

Temporal variations in microcirculatory blood flow in the testis and blood pressure were examined in intact, pentobarbital-anesthetized rats with a two-channel laser Doppler flowmeter. The laser Doppler probes that measure local blood flow in a tissue volume of about 2 mm3 were placed either over the mid portion of the left and right testes or on the right testes 1 cm apart. Testicular microcirculation was characterized by a prominent vasomotion with a frequency of 5.3+/-1.4 cycles per minute and with an amplitude of 73+/-32% (mean +/- SD) of the mean. In addition to this large and rapid variation in local blood flow, there were also major variations from minute to minute in the average blood flow, vasomotion frequency, and vasomotion amplitude at 40 and 53 minutes. Such variations in local blood flow, vasomotion frequency, and vasomotion amplitude were correlated with each other at two different sites on the same testis (r(s) = 0.39, r(s) = 0.82, r(s) = 0.64, respectively, P < 0.001), and they were all correlated with systemic blood pressure (r(s) 0.41, r(s) = 0.61, r(s) = 0.32, respectively, P < 0.001). Minute-to-minute variations in local blood flow, vasomotion frequency, and vasomotion amplitude were also correlated between the right and left testes (r(s) = 0.58, r(s) = 0.75, r(s) = 0.57, respectively, P < 0.001). There are substantial temporal variations in testicular microcirculation. These variations are to some extent related to temporal changes in systemic blood pressure, but changes in the ultralocal environment are probably more important. The functional significance of, and the factors responsible for, local variations in testicular microcirculation remain to be elucidated.

Pituitary adenylate cyclase-activating polypeptide (PACAP): effects on blood flow in the testis and caput epididymidis of the rat.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is synthesized in developing germ cells in the testis and may act as a paracrine modulator of spermatogenesis and/or participate in tubule-interstitial interactions. Despite the abundance of PACAP in the organ, its role in testicular function has not yet been studied in vivo. Using laser Doppler flowmetry, the effects of PACAP on blood flow in the testis and caput epididymidis were studied on anesthetized adult rats. When given intratesticularly as 5- and 50-ng doses, PACAP increased blood flow by 55+/-21% (mean +/- SEM, P < 0.05) and by 68+/-11% at 5 mm from the injection site, respectively. Whereas 5 ng PACAP did not influence blood flow 15 mm from the site of injection, flow was reduced (-7+/-3; P < 0.05) at this site following treatment with 50 ng. Injection of 50 ng PACAP into the caput epididymidis increased epididymal blood flow by 18+/-4% (P < 0.05) at 1 mm from the injection site. None of the treatments above significantly affected the mean arterial blood pressure. Using immunohistochemistry, PACAP was observed in elongated spermatids and in the acrosomes of round spermatids in some, but not all, seminiferous tubules. Also, distinct PACAP immunoreactivity was seen in epithelial cells, particularly in clear cells, of the caput epididymidis. In conclusion, PACAP can induce vasodilatation in both testicular and epididymal microvessels and may be involved in regulating blood flow in these organs. Whereas the vasodilatory effect of PACAP is strong in the testis, the epididymal response appears to be more moderate.

Unilateral injection of neuropeptide Y decreases blood flow in the injected testis but may also increase blood flow in the contralateral testis.

Neuropeptide Y (NPY) receptors have recently been described in intratesticular arterioles, but the role of NPY in testicular blood-flow regulation has not been examined. To explore this, we administered NPY in various doses (0.01-10 microg) via intratesticular injections and studied testicular microcirculation using a laser Doppler flow meter. NPY injection shows a dose-response pattern, with 1 microg (the most potent dose) causing a decrease (-42.4 +/- 3.7%, P < 0.00005) in blood flow in the ipsilateral testis of all the animals and an increase in blood flow in the contralateral testis (+17.2 +/- 5.6%, P = 0.03, n = 25 animals). The response in the contralateral testis was variable. A clear-cut increase was seen in 19 of the 25 animals examined, whereas either no response or a slight decrease was seen in the remaining six. The contralateral increase, which was not seen in the hindpaw on the same side, did not occur when the neuronal connections to the testes were blocked by injection of local anesthetics into the spermatic cord, either on the NPY-injected side or on the contralateral side. Our results suggest that NPY may serve as a vasoconstrictor in the testis, probably by acting on the NPY-Y1 receptors present on intratesticular arterioles. Local injection of NPY causes a major decrease in blood flow in the injected testis. This decrease is followed in the majority of animals studied by an increase in blood flow in the contralateral testis, an effect that seems to depend on neuronal mechanisms. This observation suggests that the testes may communicate under certain situations. The functional consequences of this remain to be elucidated.

Localization and effects of calcitonin gene-related peptide in the testicular vasculature of the rat.

UNLABELLED: Using laser Doppler flowmetry, the effects of unilateral intratesticular injection of calcitonin gene-related peptide (CGRP) and CGRP8-37, a CGRP-receptor antagonist, on right- and left-testicular blood flow and mean arterial pressure were studied on anesthetized adult rats. Calcitonin gene-related peptide in doses of 5 and 50 ng increased blood flow 37 +/- 11% (mean +/- SEM, P < 0.05) and 30 +/- 5% at 5 mm, but not 15 mm, away from the injection site, respectively. They did not influence mean arterial pressure nor blood flow in the contralateral testis. Five-hundred nanogram doses increased testicular blood flow in the injected testis at a point 15 mm away from the injection site (22 +/- 3%, P < 0.05) and caused a slight decrease in mean arterial pressure (-12 +/- 3%, P < 0.05). The highest dose, 5 micrograms, caused a large (-39 +/- 3%, P < 0.05) fall in mean arterial pressure within 1 minute after injection, and testicular blood flow was reduced in both the injected (-9 +/- 2%, P < 0.05, 15 mm away from injection site) and contralateral testis (-20 +/- 5%, P < 0.05). Pretreatment with 500 ng of the receptor antagonist, CGRP8-37, did not significantly attenuate the blood flow increasing affect of 50 ng CGRP, nor did 50 micrograms CGRP 8-37 (given alone) influence basal testicular blood flow in the injected testis. Using Immunohistochemistry, CGRP-containing nerves were observed in the superior and interior spermatic nerves, in the testicular artery, and in the veins leaving the testis but not in intratesticular blood vessels. CONCLUSIONS: 1) CGRP is a potent vasodilator in the testicular vasculature and it may be involved in the local regulation of testicular blood flow: 2) the testis has limited capacity to autoregulate and is consequently unable to maintain a constant testicular blood flow during large and rapid reductions in blood pressure, and 3) the local and systemic effects of vasodilators act in opposite directions in the testis.

Is nitric oxide involved in the regulation of the rat testicular vasculature?

Using immunohistochemistry, endothelial nitric oxide synthase (NOS), and neuronal NOS were localized in the endothelium of rat testicular arteries and in Leydig cells, respectively. NADPH-diaphorase activity, indicating NOS activity, however, was present only in endothelial cells. In order to examine the role of nitric oxide (NO) in the regulation of rat testicular vasculature, intact and hCG-pretreated (50-100 IU hCG given s.c. 6 h earlier) animals were given injections of the NOS inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME), 10 mg/kg i.v.). In all rats this resulted in a major increase in blood pressure. In intact, unstimulated animals, testicular vascular resistance was unaffected, and testicular blood flow consequently increased. In hCG-treated animals, in contrast, vascular resistance increased in an hCG dose-related way. L-NAME treatment also increased the hCG-induced accumulation of polymorphonuclear leukocytes in testicular venules. Treatment with N(G)-nitro-D-arginine methyl ester (D-NAME, 10 mg/kg i.v.), an inactive isomer of L-NAME, had no effect on the testicular vasculature. The study suggests that NO plays only a limited role in the regulation of testicular blood flow under basal conditions. After hCG treatment, however, NOS activity appears to be increased (increased endothelial NADPH-diaphorase staining), suggesting that NO in this situation is of importance to increase blood flow and to inhibit leukocyte accumulation.

Atrial natriuretic peptide, brain natriuretic peptide and c-type natriuretic peptide: effects on testicular microcirculation and immunohistochemical localization.

The effect of local injection of atrial (ANP), brain (BNP) and C-type (CNP) natriuretic peptides and an ANP antagonist (HS-142-1) on testicular microcirculation and vasomotion was studied using laser Doppler flowmetry. The natriuretic peptides were also localized immunohistochemically within the testis. ANP, BNP-32, CNP-22 and CNP-53 all caused a dose-related increase in testicular blood flow. The effect of ANP was blocked by concomitant injection of the ANP antagonist. Immunoreactive (ir) CNP and ir BNP were found in Leydig cells whereas in ANP was observed in the seminiferous tubules. It is suggested that the natriuretic peptides could play a role in local regulation of the testicular microcirculation.

Effects of vasoactive intestinal peptide (VIP) on the testicular vasculature of the rat.

Using laser Doppler flowmetry, the effects of unilateral intratesticular injection of vasoactive intestinal peptide (VIP) on testicular blood flow and mean arterial blood pressure were studied in anaesthetized adult rats. At doses of 50 and 500 ng, VIP increased blood flow by 29 +/- 8 and by 77 +/- 25% (p < 0.05), respectively (means +/- SEM) at 5 mm, but not at 15 mm, from the injection site. Blood pressure was not significantly affected by 50 ng, but was slightly reduced (by -8 +/- 2%) after treatment with 500 ng VIP. The highest dose of VIP (5 micrograms) reduced blood pressure (by -30 +/- 9%), and decreased blood flow at 15 mm, but not at 5 mm, from the injection site. It is concluded that VIP can act as a vasodilator in the rat testis.