Immunohistochemical analysis of estrogen receptors in the urethra of sexually intact, ovariectomized, and estrogen-substituted ovariectomized sheep.

INTRODUCTION AND HYPOTHESIS: Urinary incontinence is prevalent in postmenopausal women and spayed dogs and is associated with decreased estrogen plasma concentrations. The objective of the study was to investigate the expression of estrogen receptors (ER) in the urethra of sexually intact, ovariectomized, and estrogen-substituted ovariectomized ewes. METHODS: Paraffin cross-sections from each urethral quarter were immunohistochemically analyzed. The reactivity of ER was semiquantitatively assessed employing an immunoreactive score (IRS). RESULTS: In contrast to ERß, ERα was identified in all urethral compartments; the highest IRS was detected in the epithelium of the distal urethra. The immunoreactivity and distribution of ERα did not differ among groups. Highly significant differences in ERα concentrations were observed between consecutive urethral quarters in each group. CONCLUSIONS: Neither ovariectomy nor ovariectomy and estrogen substitution seem to have a significant effect on overall urethral ERα concentration. The results demonstrate that the precise location of the investigated urethral part is crucial to the reliable evaluation or possible comparison of ERα concentrations.

Immunohistochemical analysis of collagen types I, III, IV and alpha-actin in the urethra of sexually intact and ovariectomized beagles.

Urinary incontinence is a widespread problem in both postmenopausal women and ovariectomized dogs. The objective of this study was to investigate the influence of ovariectomy on the immunoreactivity and the distribution pattern of collagens I, III, IV and alpha-actin in the canine urethra. The immunohistochemical results were evaluated in five sexually intact and five ovariectomized beagles. The immunostaining of both collagens I and III delineated urethral connective tissue fibres and co-localized within in the fibres of both groups. The basement membranes of smooth muscle cells and sinusoids showed marked type IV collagen expression, whereas only faint immunoreactivity was present at the urothelial-stromal interface. No differences could be detected in the expression or distribution of the assessed collagen types and actin between ovariectomized and control animals. In conclusion, ovariectomy does not appear to have an effect on urethral collagens I, III, IV and smooth muscle actin in the dog, as ascertained by immunohistochemistry.

Histochemical and stereological analysis of the levator ani (pubocaudal) muscle in nulliparous and multiparous beagles.

Fiber-type composition and several stereological parameters of the levator ani (pubocaudal) muscle were evaluated in five nulliparous and five multiparous beagles using myosin ATPase-histochemistry and systematically selected muscle cross-sections. With respect to the narrow canine pelvic cavity, this study was also undertaken to determine whether vaginal birth of at least seven litters causes similar neuromuscular changes in the canine levator ani (pubocaudal) muscle analoguous to those seen in the pelvic floor muscles of women after vaginal delivery. The canine pubocaudal muscle is comprised of approximately equal amounts of slow twitch type I and fast twitch type II (IIA, IIS) fibers. The muscles of both the nulliparous and multiparous beagles did not display any signs indicative of denervation or myopathology. The multiparous dogs exhibited significantly increased mean absolute muscle (1720 mm(3)) and total fiber-type I volumes (850 mm(3)) as well as relevantly increased mean diameter of type I fibers (72.0 microm) when compared with the nulliparous group. The canine levator ani (pubocaudal) muscle is not pathologically affected by vaginal deliveries and seems to adapt to numerous successive pregnancies and births through fiber-type I hypertrophy.

Histomorphologic and morphometric evaluation of the uterine horns in nulliparous and multiparous Beagles.

OBJECTIVE: To macroscopically, histomorphologically, and morphometrically compare uterine tissues obtained from nulliparous and multiparous dogs. ANIMALS: 12 dogs constituting 2 homogenous groups (7 nulliparous Beagles and 5 multiparous Beagles, each of which had at least 7 pregnancies). PROCEDURE: Serum concentrations of progesterone and estradiol-17beta were determined. Samples of uterine tissues were fixed, embedded in paraffin, and cut into serial cross sections. Morphometric analysis was performed on systematically selected trichrome-stained sections. RESULTS: Mean absolute volume of the uterine wall did not differ between groups. Volume of blood vessels was significantly higher and relative mean value for myometrial connective tissue was significantly lower in the multiparous dogs. Arteries in the myometrium of multiparous dogs had pronounced thickening of the intima (ie, pregnancy sclerosis) and a concomitant thinning of the smooth muscle layer (tunica media). Furthermore, the elastica interna of these arteries appeared to be disintegrating and was highly and irregularly folded. Adventitia of the myometrial veins in multiparous dogs contained numerous layers of elastic fibers; however, only a few layers were observed in the adventitia of corresponding veins in nulliparous dogs. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, we documented that in contrast to other species, the uterus of nonpregnant dogs does not increase in size and volume even after at least 7 pregnancies. Furthermore, sclerotic alterations of uterine arteries are proof of at least 1 pregnancy. Results of this study may be useful in the evaluation of uterine diseases.

Differences of morphometrical parameters in hind limb muscle fibres between ovarectomized and sexually intact female dogs.

Slow and fast twitch fibres of the Mm. tibialis cranialis, semitendinosus and sartorius of seven sexually intact and seven ovarectomized female beagles were histochemically and morphometrically analysed. Along with type I and type IIA fibres, another main type II fibre (IIS), which seems to be peculiar to the dog, was found in the Mm. semitendinosus and tibialis cranialis. Type I fibers comprised 26% and type II fibres 74% of all recorded muscle fibres in the M. tibialis cranialis, 29% (type I) and 71% (type II) in the M. semitendinosus and 51% (type I) and 49% (type II) in the M. sartorius, respectively. The average single profile area and the corresponding mean diameter of fibre types I and II in the investigated hind limb muscles were generally larger in ovarectomized than in sexually intact animals. This was more evident in type II than in type I fibres. However only the type II fibres of the M. tibialis cranialis and sartorius exhibited a statistically significant increase in diameter (p < 0.01 and p < 0.05, respectively). Accordingly, the mean density (number of fibres/mm2) of both fibre types in the hind limb muscles of spayed dogs was generally reduced. Again, this reduction attained statistical relevance in the type I and II fibres of the tibialis cranialis. In addition, the fibre densities of type I in the semitendinosus and type II in the sartorius muscles were also significantly reduced in ovarectomized dogs. In conclusion, ovarectomized beagles showed a generally increased mean diameter of the investigated type I and II hind limb muscle fibres and a concomitant decreased average fibre density of the respective types when compared to sexually intact animals.