A Cytochemical Analysis of the Lactic Dehydrogenase-Diphosphopyridine Nucleotide-Diaphorase System in the Epididymis of the Mouse

Abstract

The properties of diphosphopyridine nucleotide diaphorase and the lactic dehydrogenase-diphosphopyridine nucleotide diaphorase system in the epididymis of the mouse have been investigated by cytochemical and quantitative methods. In normal mice diphosphopyridine nucleotide diaphorase activity was homogeneously distributed within the cytoplasm of the epithelial cells of the epididymal canal. A gradient of activity existed along the duct, with the lowest levels of activity appearing in the head segments and the highest levels in the body and tail segments. The lactic dehydrogenase-diphosphopyridine nucleotide diaphorase system differed in its cytological distribution from diphosphopyridine nucleotide diaphorase. In the cells of the body portion of the epididymal canal this system showed high levels of activity in the cell apex with low levels of activity prevailing in the general cytoplasm. In other areas the system showed low levels of activity homogeneously distributed throughout the cytoplasm. Cytochemical studies indicated that the activity of diphosphopyridine nucleotide diaphorase was depressed by orchidectomy and restored by testosterone propionate administration. The activity of this enzyme was unaffected by section of the vasa efferentia (vasectomy). The activity of the lactic dehydrogenase-diphosphopyridine nucleotide diaphorase system was depressed by vasectomy and orchidectomy. Testosterone propionate restored activity of tissues from castrated animals to levels prevailing in vasectomized animals but did not re-establish the distribution pattern observed in normal animals. Quantitative determinations confirmed and extended cytochemical observations.

On the regional histology and histochemistry of the epididymis of the camel (Camelus dromedarius)

“…Its activity was highest in the terminal segment, confirming the findings of Erkmann (1971) that the highest activity of LDH, G-6-PDH and SDH was located in the head and tail segments of the bull epididymis although Stallcup & Roussel (1965) reported the highest activity in the corpus epididymidis. In the mouse the lowest levels of LDH activity were found in the head segment and the highest in the body and tail segments (Allen & Slater, 1961 …”

“…Regional differences based on histological and/or histochemical observations have been reported by Montagna (1952), Cavazos (1956), Allen & Slater (1957, 1958, 1961, Nicander (1957Nicander ( , 1958, Reid & Cleland (1957), Blackshaw & Samisoni (1967), Linnetz & Amann (1968), Erkmann (1971), Moniem (1972b) and Moniem & Glover (1972). Studies of the functional activity of the lining epithelia have included absorption of vital dyes such as trypan blue and India ink (Mason & Shaver, 1952;Shaver, 1954;Macmillan, 1957;Macmillan & Clegg, 1963;Burgos, 1964Burgos, , 1967Sedar, 1966;Friend & Farquhar, 1967), incorporation of isotopes (Orgebin-Crist, 1964) and sperm and fluid résorption under normal and experimental conditions (Crabo & Gustafsson, 1964;Crabo, 1965;Nicander, 1965;Holstein, 1967;Hoffer, Hamilton & Fawcett, 1973).…”

Cytochemical Localization of Lactic Dehydrogenase in White Skeletal Muscle

“…Nachlas et al (16,40) and Hess et al (17), who described the histochemical methods which are now in wide use for coenzymelinked dehydrogenases, presumed that the diaphorases were distributed abundantly in all tissues in association with dehydrogenases. Recently, however, Allen and Slater reported (20) that in the mouse epididymis the L D H system gave a weaker reaction histochemically than did the muscle stained much more strongly for L D H than did the white muscle, although biochemical assays of tissue homogenates indicated that white muscle had higher levels of L D H activity than did the red muscle (41). At the time of preparation of this manuscript, these authors reported also (22) that the addition of PMS to the staining system for L D H increased the staining of the white muscle fibers.…”

“…H + + pyruvate + NADH LDH , Lactate + NAD + methods for LDH (16,17) is limited by (a) dependence of the staining results on the presence in tissue of a diaphorase which must be associated with the dehydrogenase to transfer the electrons from the reduced coenzyme I(NADH) to the tetrazofium salt (18)(19)(20)(21)(22) and (b) by the leakage of LDH from tissue sections into the incubation medium (22)(23)(24)88). In this study, (a) the limitations of the standard histochemical methods for LDH are analyzed quantitatively; (b) evidence is presented that the standard methods are inadequate to reflect the sites of enzymatic activity in white skeletal muscle; (c) a modified method for cytochemical localization of LDH is presented which prevents the diffusion of LDH into the incubation medium and, at the same time, makes the staining system independent of tissue diaphorase; and (d) it is demonstrated that this modified method when applied to the adductor magnus muscle of the rabbit reveals a fine reticulum in the sarcoplasm of muscle fibers in addition to the usual staining of the mitochondria.…”

Comparative histochemical localization of some hydrolytic enzymes in mammalian epididymides

“…A supplementary control for alkaline phospha tase was incubated in a similar medium that contained 4 mm M-cysteine hydrochloride in the form of powder, prior to the adjustment of pH. Similar concentrations of cysteine hydrochloride were also added to substrates prepared for demonstration of adenosine triphospha tase and adenosine monophosphatase to inhibit the alkaline phosphatase activity [Allen and Slater, 1961).…”