Optimal Wire Myography Normalization for the Rat Dorsal Penile, Internal Pudendal and Internal Iliac Arteries

Abstract

In functional arterial studies using wire myography, the determination of a vessel’s standardized normalization factor (factor k) is an essential step to ensure optimal contraction and relaxation by the arteries when stimulated with their respective vasoactive agents and to obtain reproducible results. The optimal factor k for several arteries have been determined, however, the optimal initial tension and factor k for the arteries involved in erection remains unknown. Hence, in the present study we set out to determine the optimal factor k for the internal iliac artery, proximal and distal internal pudendal artery (IPA), and dorsal penile artery. After isolating, harvesting, and mounting the arteries from male Sprague-Dawley rats on a multi wire myograph, we tested arterial responsivity to high K+-stimulation when the factor k was set at 0.7, 0.8, 0.85, 0.9, 0.95, 1.0, 1.1, and 1.2 to determine the factor k setting that results in the greatest K+-induced active force production for each vessel type. The data showed the optimal factor k is 0.90-0.95 for the dorsal penile, distal internal pudendal and internal iliac arteries while it is 0.85-0.90 for proximal internal pudendal artery. These optimal values corresponded to initial passive tension settings of 1.10±0.16 - 1.46±0.23, 1.28±0.20 - 1.69±0.34, 1.03±0.27 - 1.33±0.31, and 1.33±0.31 - 1.77±0.43 mN/mm for the dorsal penile, distal IP, proximal IP, and internal iliac arteries, respectively.

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“…Penile tissue was exposed by freeing the tissue of skin and fascia, and penile tissue from the base to the proximal glans was harvested. The distal segment of the internal pudendal artery (dIPA) was obtained by dissection from 1 mm distal to the gluteal artery bifurcation to the point where the IPA terminates by dividing into the dorsal and deep penile arteries, as described in further detail 54 . The vessels and the penile tissues were further cleaned of connective tissue under a dissection microscope in ice‐cold Kreb's buffer (NaCl 130 mM, KCl 4.7 mM, K 3 PO 4 1.18 mM, Mg 2 SO 4 1.18 mM, Na 2 CO 3 14.9 mM, D‐Glucose 5.6 mM, CaCl 2 1.56 mM and EDTA 0.03 mM).…”

“…The distal segment of the internal pudendal artery (dIPA) was obtained by dissection from 1 mm distal to the gluteal artery bifurcation to the point where the IPA terminates by dividing into the dorsal and deep penile arteries, as described in further detail. 54 The vessels and the penile tissues were further cleaned of connective tissue under a dissection microscope in ice-cold Kreb's buffer (NaCl 130 mM, KCl 4.7 mM, K 3 PO 4 1.18 mM, Mg 2 SO 4 1.18 mM, Na 2 CO 3 14.9 mM, D-Glucose 5.6 mM, CaCl 2 1.56 mM and EDTA 0.03 mM). The dorsal vein, corpus spongiosum, and connective and adventitial tissues were carefully excised from the penile shaft using a micro spring scissors.…”