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. 2001 Mar;132(6):1333-41.
doi: 10.1038/sj.bjp.0703938.

Morphological and functional in vitro and in vivo characterization of the mouse corpus cavernosum

H Mizusawa  1 , P Hedlund, A Håkansson, P Alm, K E Andersson
Affiliations

Morphological and functional in vitro and in vivo characterization of the mouse corpus cavernosum

H Mizusawa et al. Br J Pharmacol. 2001 Mar.

Abstract

1. In normal mice, the distribution of adrenergic, cholinergic, some peptidergic, and neuronal nitric oxide synthase (nNOS)-containing nerves were investigated. Functional in vitro correlates were obtained. An in vivo model was developed in which erectile haemodynamics in response to drugs or nerve-stimulation were studied. 2. Immunoreactivities for vesicular acetylcholine transporter protein (VAChT), nNOS-, and vasoactive intestinal polypeptide (VIP), co-existed in nerve fibres and terminal varicosities. Immunoreactivities for neuropeptide Y (NPY) and tyrosine hydroxylase (TH) were found in the same nerve structures. 3. Chemical sympathectomy abolished TH- and NPY-IR nerve structures in cavernous smooth muscle bundles. The distribution of calcitonin gene-related peptide (CGRP)-, nNOS-, VAChT- and VIP-IR nerve structures was unchanged. 4. In endothelial cells of the central and helicine arteries, veins and venules, intense immunoreactivity for endothelial NOS (eNOS) was observed. No distinct eNOS-IR cells were found lining the cavernous sinusoids. 5. In vitro, nerve-induced relaxations were verified, and endothelial NO/cyclic GMP-mediated relaxant responses were established. VIP and CGRP had small relaxant effects. A functioning adenylate cyclase/cyclic AMP pathway was confirmed. 6. Neuronal excitatory responses were abolished by prazosin, or forskolin. VIP and CGRP counteracted contractions, whereas NPY and scopolamine enhanced excitatory responses. 7. In vivo, erectile responses were significantly attenuated by L-NAME (50 mg kg(-1)) and facilitated by sildenafil (200 microg kg(-1)). 8. It is concluded that the mouse is a suitable model for studies of erectile mechanisms in vitro and in vivo.

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Figures

Figure 1
Figure 1
The mouse penis and technical landmarks for the registration of intracavernous pressure in vivo.
Figure 2
Figure 2
Typical curve showing the increase in intracavernous pressure (ICP) induced by nerve stimulation in vivo. During stimulation, the time for ICP to reach 80% of maximal increase (peak ICP – basal ICP) was recorded (T80). At this point, the increase per second (ΔT80) was evaluated. After stimulation, the time for and the rate by which a decrease to 20% of maximal pressure occurred (D20 and ΔD20) were determined.
Figure 3
Figure 3
Mouse corpus cavernosum. Double immunohistochemistry. (a) Vesicular acetylcholine transporter (VAChT)-immunoreactive (IR) nerve terminals along smooth muscle bundles. FITC immunofluorescence. (b) Same section as in (a). NO synthase-IR nerve terminals displaying coinciding profiles with VAChT-IR varicosities. (c) Tyrosine hydroxylase (TH)-IR nerve terminals. FITC immunofluorescence. (d) Neuropeptide Y-IR nerve terminals exhibiting coinciding profiles with TH-IR nerve terminals. Same section as in (d). Texas red immunofluorescence. Bars=100 μm
Figure 4
Figure 4
Mouse corpus cavernosum. Triple immunocytochemistry. Confocal microscopy. (a) Vesicular acetylcholine transporter – immunoreactive (IR) nerve terminals. FITC- immunofluorescence. (b) Same section as in (a). NO synthase-IR nerve varicosities. Texas red immunofluorescence. (c) Same section as in (a) and (b). Vasoactive intestinal polypeptide-containing varicosities. Cy-5 immunofluorescence. Each image consists of 19 consecutive sections with 0.30 μm between adjacent sections. Bars=10 μm.
Figure 5
Figure 5
Tracing showing the effects of forskolin on electrical field stimulation-induced contractions in an isolated preparation of mouse corpus cavernosum (top) and on intracavernous pressure in an anaesthetized mouse after intracavernous administration (bottom).
Figure 6
Figure 6
Intracavernous pressure (ICP) changes in the mouse corpus cavernosum in response to cavernous nerve stimulation in vivo. After pretreatment with intraperitoneally administered L-NAME (50 mg kg−1), the erectile response was abolished. The increase in ICP induced by forskolin (5 μg kg−1) was unaffected by L-NAME.
Figure 7
Figure 7
Intracavernous pressure (ICP) changes in the mouse corpus cavernosum in response to cavernous nerve stimulation (3 V, 10 Hz) in vivo. Facilitatory effect of sildenafil (200 μg kg−1).
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