The specificity of the sialidase activity present in rat kidney cortex (12 000 × g pellet) was studied with various tritiated oligosaccharidic substrates: (i) αNeuAc2 → 3βGall → 4Glc‐itol[3H], αNeuAc2 → 6βGall → 4Glc‐itol[3H] and αNeuAc2 → 8αNeuAc2 → 3βGall → 4Glc‐itol[3H] from bovine colostrum; (ii) α‐NeuAc2 → 6βGall → 4βGlcNAc‐itol[3H], αNeuAc2 → 3βGal1 → 4βGlcNAcl → 2αManl → 3βMan1 → 4GlcNAc‐itol[3H]. αNeuAc2 → 6βGall → 4βGlcNAcl → 2αManl α 3(βGall → 4GlcNAcl → 2αManl → 6)βManl → 4GlcNAc‐itol [3H]et αNeuAc2 → 6βGall → 4βGlcNAcl → 2αManl‐3(αNeuAc2 → 6βGall → 4βGlcNAcl → 2αManl → 6)βManl 4GlNAc‐itol[3H] isolated from the urine of a patient with mucolipidosis I. The enzyme cleaves α2 → 3 and α2 → 8 linkages at a greater rate than the α2 → 6 bonds. Its activity decreases with the length of the oligosaccharidic chain. Substitution of a glucose moiety by Nacetylglucosamine results in diminished activity. The specificity of rat kidney sialidase differs from that reported for other mammalian of viral sialidases.
Because kidney microangiopathy with capillary basement membrane thickening has been reported in spontaneous hypertension, we have studied the activities of three lysosomal glycosidases able to degrade the carbohydrate moieties of basement membrane constituents in the kidney cortex of 12-week-old spontaneously hypertensive rats (SHR) and age-matched normotensive Wistar Kyoto rats (WKY). These activities were also determined in SHR and WKY treated from 6 to 12 weeks of age with hydralazine (mean dose, 18 mg/kg per day in drinking water). Sialidase specific activity on sialyl-alpha 2-3-[3H]lactitol was markedly decreased in the kidney of untreated SHR, 40% activity remaining relative to that found in untreated age-matched WKY (p less than 0.001). beta-Galactosidase specific activity on p-nitrophenyl-beta-D-galactoside was also decreased, 86% activity remaining relative to that found in untreated WKY (p less than 0.001). Glucosyl-galactosyl-hydroxylysyl glucohydrolase specific activity on glucosyl-galactosyl-hydroxylysine was equally diminished, 74% activity remaining relative to that found in untreated age-matched WKY (p less than 0.001). In contrast, the activities of two control glycosidases inactive on the carbohydrate moieties of basement membrane constituents, alpha-glucosidase assayed with p-nitrophenyl-alpha-D-glucoside as substrate and beta-glucosidase assayed with p-nitrophenyl-beta-D-glucoside as substrate, were significantly increased. All the alterations in enzyme activities observed in the kidney of SHR were also present in the long-term treated normotensive SHR. No effect of the hydralazine treatment on the three enzyme activities investigated could be demonstrated in the WKY. Thus the alterations observed in the kidneys of SHR appear to be independent of blood pressure level.
Rat kidney sialidase levels have been reported to be markedly altered in pathological
states such as diabetes. This was associated with a modification of sialic acid levels.
Therefore, it was interesting to study the variations of kidney sialidase and sialyltransferase
activities and sialic acid content according to sex and age. This was carried out from birth to
210 days of age. The substrates used were sialyla(2-3)[^3H]-lactitol for sialidase activity, asialofetuin
and [^14C]-CMPNeu5Ac for sialyltransferase activity. In males sialidase activity
increased until 32 days then slightly declined. In females, the activity increased and leveled
off at 135 days of age. Higher sialidase activity was observed in females than in males from
56 days of age. Gonadectomy had no effect on this activity. In both sexes, sialyltransferase
activity decreased markedly with age. This activity was higher in females than in males,
whereas sialic acid levels varied only moderately with age and were slightly higher in
females.
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