THE HISTOCHEMISTRY OF THE TESTIS TISSUES FROM PATIENTS OF IDIOPATHIC MALE INFERTILITY

Report I. Studies on Complex Carbohydrate (Localization of Acid Mucopolysaccharides)

Abstract

In the present study, testis tissues were obtained from patients (in 23 cases) of idiopathic male infertility whose semen specimens were diagnosed to be either oligozoospermia or azoospermia. Testis tissues were furthermore obtained from patients (in 4 cases) of cryptorchism, eunuchoidism, bilateral congenital absence of the vas deferens and postvascectomized azoospermia. At the time of each surgical procedure, control specimens were dissected out from 6 individuals who were not clinically suffering from idiopathic male infertility. In all cases, Charny's accepted biopsy technique was employed. The tissue specimens were subjected to the following three types of fixation, 1) Carnoy's fluid (for 2 hours at room temperature), 2) 1% cetylpyridinium chloride (cpc) in Carnoy's fluid (for 2 hours at room temperature) and 3) 1% cpc in Carnoy's fluid (for 2 hours at room temperature) following treatment with cpc-urea for 2 hours at 53°C. After fixation, all the tissue specimens were embedded in paraffin and then stained with hematoxylin-eosin for general observation as well as the following histochemical methods for the demonstration of complex carbohydrates: 1) alcian blue (AB) (pH 1.0), 2) AB (pH 2.5), 3) dialyzed iron ferrocyanide (DI-FCY), 4) periodic acid-Schiff (PAS). For identifying individual acidic and neutral carbohydrates, the following procedures of enzyme digestion were performed upon some tissue sections prior to histochemical staining, a) sialidase [prior to staining with AB (pH 2.5)], b) streptomyces hyaluronidase [prior to staining with AB (pH 2.5)], c) testicular hyaluronidase [prior to staining with AB (pH 1.0) or (pH 2.5)], d) chondroitinase ABC [prior to staining with AB (pH 1.0) or pH 2.5)], e) chondroitinase AC [prior to staining with AB (pH 1.0) or (pH 2.5)], f) α-amylase [prior to staining with PAS].
The results are summarized as follows:
1) The seminiferous tubular walls and interstitial connective tissue in the testis tissues from both the normal and infertile individuals were believed to contain acidic complex carbohydrates with sulfate esters and carboxyl groups.
2) The present effects of digestion with chondroitinase ABC and AC upon AB (pH 1.0 and 2.5) reactions of the seminiferous tubular walls in the testis tissues from normal individuals and patients of male infertility were taken to indicate the presence of isomeric chondroitin sulfates (A and/or C) in these structure.
3) The interstitium of the testis tissues from normal individuals and patients of male infertility contained chondroitin sulfate B.
4) According to the present results obtained by the PAS staining procedure employed in combination with digestion with α-amylase, the germinal and supporting cells in the infertile testis tissues were thought to contain a large amount of glycogen than those in the cells in normal testis tissues.