Vogel P, Payne B J, Read R, Lee W-S, Gelfman C M, Kornfeld S
Lexicon Pharmaceuticals, Pathology Department, 8800 Technology Forest Place, The Woodlands, TX 77381-1160, USA.
Vet Pathol. 2009 Mar;46(2):313-24. doi: 10.1354/vp.46-2-313.
UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine-1-phosphotransferase (GlcNAc-1-phosphotransferase) is an alpha(2)beta(2)gamma(2) hexameric enzyme that catalyzes the first step in the synthesis of the mannose 6-phosphate targeting signal on lysosomal hydrolases. In humans, mutations in the gene encoding the alpha/beta subunit precursor give rise to mucolipidosis II (MLII), whereas mutations in the gene encoding the gamma subunit cause the less severe mucolipidosis IIIC (MLIIIC). In this study we describe the phenotypic, histologic, and serum lysosomal enzyme abnormalities in knockout mice lacking the gamma subunit and compare these findings to those of mice lacking the alpha/beta subunits and humans with MLII and MLIIIC. We found that both lines of mutant mice had elevated levels of serum lysosomal enzymes and cytoplasmic alterations in secretory cells of several exocrine glands; however, lesions in gamma-subunit deficient (Gnptg(-/-)) mice were milder and more restricted in distribution than in alpha/beta-subunit deficient (Gnptab(-/-)) mice. We found that onset, extent, and severity of lesions that developed in these two different knockouts correlated with measured lysosomal enzyme activity; with a more rapid, widespread, and severe storage disease phenotype developing in Gnptab(-/-) mice. In contrast to mice deficient in the alpha/beta subunits, the mice lacking the gamma subunits were of normal size, lacked cartilage defects, and did not develop retinal degeneration. The milder disease in the gamma-subunit deficient mice correlated with residual synthesis of the mannose 6-phosphate recognition marker. Of significance, neither strain of mutant mice developed cytoplasmic vacuolar inclusions in fibrocytes or mesenchymal cells (I-cells), the characteristic lesion associated with the prominent skeletal and connective tissue abnormalities in humans with MLII and MLIII. Instead, the predominant lesions in both lines of mice were found in the secretory epithelial cells of several exocrine glands, including the pancreas, and the parotid, submandibular salivary, nasal, lacrimal, bulbourethral, and gastric glands. The absence of retinal and chondrocyte lesions in Gnptg(-/-) mice might be attributed to residual beta-glucuronidase activity. We conclude that mice lacking either alpha/beta or gamma subunits displayed clinical and pathologic features that differed substantially from those reported in humans having mutations in orthologous genes.
UDP-N-乙酰葡糖胺:溶酶体酶N-乙酰葡糖胺-1-磷酸转移酶(GlcNAc-1-磷酸转移酶)是一种α(2)β(2)γ(2)六聚体酶,催化溶酶体水解酶上甘露糖6-磷酸靶向信号合成的第一步。在人类中,编码α/β亚基前体的基因突变会导致黏脂贮积症II型(MLII),而编码γ亚基的基因突变会导致病情较轻的黏脂贮积症IIIC型(MLIIIC)。在本研究中,我们描述了缺乏γ亚基的基因敲除小鼠的表型、组织学和血清溶酶体酶异常情况,并将这些发现与缺乏α/β亚基的小鼠以及患有MLII和MLIIIC的人类的情况进行比较。我们发现,这两种突变小鼠品系的血清溶酶体酶水平均升高,且几个外分泌腺的分泌细胞出现细胞质改变;然而,γ亚基缺陷(Gnptg(-/-))小鼠的病变比α/β亚基缺陷(Gnptab(-/-))小鼠更轻微,分布也更局限。我们发现,这两种不同基因敲除小鼠中出现的病变的起始、范围和严重程度与所测的溶酶体酶活性相关;Gnptab(-/-)小鼠出现了更迅速、广泛且严重的贮积病表型。与缺乏α/β亚基的小鼠不同,缺乏γ亚基的小鼠体型正常,没有软骨缺陷,也未发生视网膜变性。γ亚基缺陷小鼠病情较轻与甘露糖6-磷酸识别标记的残留合成有关。重要的是,这两种突变小鼠品系均未在成纤维细胞或间充质细胞(I细胞)中出现细胞质空泡内含物,而这种特征性病变与患有MLII和MLIII的人类明显的骨骼和结缔组织异常相关。相反,这两种小鼠品系的主要病变出现在几个外分泌腺的分泌上皮细胞中,包括胰腺以及腮腺、颌下唾液腺、鼻腺、泪腺、尿道球腺和胃腺。Gnptg(-/-)小鼠没有视网膜和软骨细胞病变可能归因于残留的β-葡糖醛酸酶活性。我们得出结论,缺乏α/β或γ亚基的小鼠表现出的临床和病理特征与在直系同源基因突变的人类中报道的情况有很大不同。