Significant Muscle Capillary Basement Membrane Thickening in Spontaneously Diabetic Mystromys Albicaudatus Yohannes W. Yesus, M.D., James A. Esterly, M.D., Robert A. Stuhlman, D.V.M., M.S., and John F. Townsend, M.D., Columbia, Mo.
SUMMARY
Skeletal muscle capillary basement membrane thickness was determined in age- and sex-matched normal and spontaneously diabetic Mystromys albicaudatus. In nondiabetic animals the average basement membrane thickness was 482.6 ± 48.7 A as against 779.0 ± 319.9 A in the diabetic ones. This difference was statistically significant at P23 ' 24 None of these animal models shows the wide range of signs and diverse pathology that has been described in man. Particularly lacking in animal models is the almost universal finding in human diabetics of thickened muscle capillary basement membranes. Bloodworth et al. 1 5 reported significant glomerular, retinal, and peripheral capillary basement membrane thickening in dogs made long-term diabetics by alloxan or bovine growth hormone. However, data from similar attempts in rats and rabbits have been inconclusive. 16 Salazar et al. 1 7 reported the absence of ultrastructural muscle capillary basement membrane change in streptozotocin-induced diabetes in the rhesus monkey. Moreover, the use of models with induced diabetes will not allow the analysis of the nature and the pathogenic importance of hereditary components that may be important factors leading to the occurrence of microvascular lesions. In other spontaneously diabetic animals, including the Chinese hamster, no microvascular changes comparable to the muscle capillary basement membrane changes in human diabetics have been reported. In this preliminary report on .the animal model M. al448
bicaudatus, the number of animals was limited because only animals that were age- and sex-matched and severely diabetic were included. The number of readily available animals that fulfill these criteria is limited, and long waiting periods are necessary to obtain large numbers of severe diabetics, unless large breeding colonies are established. However, even with the relatively small number of animals, the difference in muscle capillary basement membrane thickness between diabetics and nondiabetics is highly significant. On the basis of 95 per cent confidence limits, 75 per cent prevalence of significant basement membrane thickening was noted in the animals studied as against 56 per cent in the report of Williamson et al. 5 The lack of difference in AMBMT in outside- and inside-colony nondiabetics indicated that the basement membrane changes are not the result of local colony environmental factors but instead are related to the genetically determined carbohydrate intolerance. The pattern of basement membrane thickening in this animal model appears to be similar to that described in human diabetics. Williamson et al. 2 5 have used the S.D. of the AMBMT in each individual to represent the segmental nature of basement membrane change in diabetes. They have found significant difference in S.D.s in diabetics as compared with normals. Similar change is demonstrated in these diabetic animals. Some of the markedly thickened capillary basement membranes from diabetic animals showed a tendency toward reduplication of the basement membrane. This finding has been stressed by Vracko et al. 2 6 in their study of human diabetics. The interesting and crucial question of relationship of duration of diabetes to basement membrane thickness is raised in this study. In this limited number of animals a high positive correlation is demonstrated. Further extension of the study to large numbers of animals is necessary to confirm this preliminary observation. This animal model stands apart from other spontaneously diabetic animals in several important aspects of the disease. First, it is the only model reported to show thickened muscle capillary basement membranes. Secondly, renal glomerular changes, including increased mesangial areas and thickened glomerular capillaries, are seen in diabetic Mystromys, beyond the glomerulosclerosis due to age alone. 26 Third, there is no relationship between obesity and hyperglycemia. 1819 These manifestations of the diabetic syndrome in this species closely resemble the manfestations of human diabetes mellitus. DIABETES, VOL. 2 5 , NO. 5
YOHANNES W. YESUS, M.D., AND ASSOCIATES
ACKNOWLEDGMENTS
The authors wish to express their appreciation to Mr. Donald Thompson and Mrs. Barbara J. Ballenger for their invaluable technical assistance with electron microscopy, Dr. David P. Hutcheson for his help in the statistical analyses of these data, and Mrs. Janice Franz for her technical assistance in all samplings and tissue harvesting. This research was supported in part by grant 510118-1 from the American Diabetes Association. REFERENCES 'Zach, S. I., Peques, J. J., and Elliot, F. A.: Interstitial muscle capillaries in patients with diabetes mellitus: A light and electron microscope study. Metabolism 77:381-93, 1962. 2 Siperstein, M. D., Norton, W., Unger R. H., and Madison, D. L.: Diabetic capillary basement membrane width in normal, diabetic and prediabetic patients. Trans. Assoc. Am. Physicians 79:330-44, 1966. 3 Siperstein, M. D., Unger, R. H., and Madison, L. L.: Studies of muscle capillary basement membrane in normal subjects, diabetic and prediabetic patients. J. Clin. Invest. 47:1973-99, 1968. 4 Williamson, J. R., and Kilo, C : Basement membrane thickening and the mystery of diabetes. Hospital Practice: .52:109-17, 1971. 5 Williamson, J. R., Vogler, N. J., and Kilo, C : Estimation of vascular basement membrane thickness. Diabetes 78:567-78, 1969. 6 Bencosme, S. A., West, R. O., Kerr, J. W., and Willson D. L.: Diabetic capillary angiopathy in human skeletal muscle. Am. J. Med. 40:67-77, 1966. 7 Kilo, C , Vogler, N., and Williamson, J. R.: Muscle capillary basement membrane changes related to aging and diabetes. Diabetes 27:881-905, 1972. 8 Danowski, T. S., Fisher, E. R., Khurana, R. C , Nolan, S., and Stephen, T.: Muscle capillary basement membranes in juvenile diabetes mellitus. Metabolism 27:1125-32, 1972. 9 Williamson, J. R., Vogler, N. J., and Kilo, C : Microvascular diseases in diabetes. Med. Clin. N. Am. .5.5:847-60, 1971. 10 Vracko, R., and Strandness, D. E.: Basal lamina of abdominal skeletal muscle capillaries in diabetics and nondiabetics. Circulation 3.5:690-700, 1967. n Vracko, R.: Skeletal muscle capillaries in diabetics: A quantitative analysis. Circulation 47:271-83, 1970.
MAY, 1976
12
Fagerberg, S. E.: Studies in pathogenesis of diabetic neuropathy. Acta Med. Scand. 7.54:145-50, 1956. 13 Aagenaes, O., and Mae, H.: Light- and electron-microscopic study of skin capillaries of diabetics. Diabetes 70:253-59, 1961. 14 Frantizis, T. G.: The ultrastructure of capillary basement membranes in the attached gingiva of diabetic and non-diabetic patients. J. Periodontol. 42:49, 1971. 15 Bloodworth, J. M. B., Jr., Engerman, R. L., and Powers, K. L.: Experimental diabetic microangiopathy: Basement membrane statistics in the dog. Diabetes 78:455-58, 196916 Cameron, D. P., Amherdt, M., Leuenberger, P., Orci, L., and Stauffacher, W.: Microvascular alterations in chronically streptozotocin-diabetic rats. In Vascular and Neurological Changes in Early Diabetes. Camerini-Davalos, R. A., and Cole, H. S., Eds. New York, Academic Press, 1973, pp. 257-64. 17 Salazar, H., Chez, R. A., and Pardo, M.: Absence of ultrastructural changes in the basement membrane of muscle capillaries in streptozocin-induced carbohydrate intolerance in rhesus monkeys. Am. J. Pathol. 77:437-45, 1973. 18 Packer, J. T., Kraner, K. L , Rose, S. D., Stuhlman, R. A., and Nelson, L. R.: Diabetes mellitus in Mystromys albicaudatus. Arch. Pathol. 89:410-15, 1970. 19 Stuhlman, R. A., Packer, J. T., and Doyle, R. E.: Spontaneous diabetes mellitus in Mystromys albicaudatus: Repeated glucose values from 620 animals. Diabetes 27:715-21, 1972. 20 Stuhlman, R. A.: The genetic mode of transmission of spontaneous diabetes mellitus in Mystromys albicaudatus: Columbia, University of Missouri, M.S. thesis, 1971. 21 Stuhlman, R. A., Srivastava, P. K., Schmidt, G., Vorbeck, M. L., and Townsend J. F.: Characterization of diabetes mellitus in South African hamsters (Mystromys albicaudatus). Diabetologia 70:685-90, 1974. 22 Stuhlman, R. A., Packer, J. T., and Rose, S. D.: Repeated blood sampling of Mystromys albicaudatus (white tailed rat). Lab. Anim. Sci. 22:268-70, 1972. 23 Renold, A. E., and Burr, I.: The pathogenesis of diabetes mellitus: Possible usefulness of spontaneous hyperglycemic syndromes in animals. Calif. Med. 7 72:23-34, 1970. 24 Renold, A. E., and Dulin, W. E., Eds.: Spontaneous diabetes in laboratory animals. Diabetologia 3.63-286, 1967. "Williamson, J. R., Vogler, N. J., and Kilo C : Early capillary basement membrane changes in subjects with diabetes mellitus. Adv. Metab. Disord. Suppl. 2:363-67, 1973. 26 Vracko, R. and Benditt, E. P.: Manifestations of diabetes mellitus—Their possible relationship to an underlying cell defect. Am. J. Pathol. 7.5:204-22, 1974. 27 Riley, T., Stuhlman, R. A., Van Peenen, H. J., Esterly, J. A., and Townsend, J. F.: Glomerular lesions of diabetes mellitus in Mystromys albicaudatus. Arch. Pathol. 99:167-69, 1975.
449
SUMMARY
Skeletal muscle capillary basement membrane thickness was determined in age- and sex-matched normal and spontaneously diabetic Mystromys albicaudatus. In nondiabetic animals the average basement membrane thickness was 482.6 ± 48.7 A as against 779.0 ± 319.9 A in the diabetic ones. This difference was statistically significant at P23 ' 24 None of these animal models shows the wide range of signs and diverse pathology that has been described in man. Particularly lacking in animal models is the almost universal finding in human diabetics of thickened muscle capillary basement membranes. Bloodworth et al. 1 5 reported significant glomerular, retinal, and peripheral capillary basement membrane thickening in dogs made long-term diabetics by alloxan or bovine growth hormone. However, data from similar attempts in rats and rabbits have been inconclusive. 16 Salazar et al. 1 7 reported the absence of ultrastructural muscle capillary basement membrane change in streptozotocin-induced diabetes in the rhesus monkey. Moreover, the use of models with induced diabetes will not allow the analysis of the nature and the pathogenic importance of hereditary components that may be important factors leading to the occurrence of microvascular lesions. In other spontaneously diabetic animals, including the Chinese hamster, no microvascular changes comparable to the muscle capillary basement membrane changes in human diabetics have been reported. In this preliminary report on .the animal model M. al448
bicaudatus, the number of animals was limited because only animals that were age- and sex-matched and severely diabetic were included. The number of readily available animals that fulfill these criteria is limited, and long waiting periods are necessary to obtain large numbers of severe diabetics, unless large breeding colonies are established. However, even with the relatively small number of animals, the difference in muscle capillary basement membrane thickness between diabetics and nondiabetics is highly significant. On the basis of 95 per cent confidence limits, 75 per cent prevalence of significant basement membrane thickening was noted in the animals studied as against 56 per cent in the report of Williamson et al. 5 The lack of difference in AMBMT in outside- and inside-colony nondiabetics indicated that the basement membrane changes are not the result of local colony environmental factors but instead are related to the genetically determined carbohydrate intolerance. The pattern of basement membrane thickening in this animal model appears to be similar to that described in human diabetics. Williamson et al. 2 5 have used the S.D. of the AMBMT in each individual to represent the segmental nature of basement membrane change in diabetes. They have found significant difference in S.D.s in diabetics as compared with normals. Similar change is demonstrated in these diabetic animals. Some of the markedly thickened capillary basement membranes from diabetic animals showed a tendency toward reduplication of the basement membrane. This finding has been stressed by Vracko et al. 2 6 in their study of human diabetics. The interesting and crucial question of relationship of duration of diabetes to basement membrane thickness is raised in this study. In this limited number of animals a high positive correlation is demonstrated. Further extension of the study to large numbers of animals is necessary to confirm this preliminary observation. This animal model stands apart from other spontaneously diabetic animals in several important aspects of the disease. First, it is the only model reported to show thickened muscle capillary basement membranes. Secondly, renal glomerular changes, including increased mesangial areas and thickened glomerular capillaries, are seen in diabetic Mystromys, beyond the glomerulosclerosis due to age alone. 26 Third, there is no relationship between obesity and hyperglycemia. 1819 These manifestations of the diabetic syndrome in this species closely resemble the manfestations of human diabetes mellitus. DIABETES, VOL. 2 5 , NO. 5
YOHANNES W. YESUS, M.D., AND ASSOCIATES
ACKNOWLEDGMENTS
The authors wish to express their appreciation to Mr. Donald Thompson and Mrs. Barbara J. Ballenger for their invaluable technical assistance with electron microscopy, Dr. David P. Hutcheson for his help in the statistical analyses of these data, and Mrs. Janice Franz for her technical assistance in all samplings and tissue harvesting. This research was supported in part by grant 510118-1 from the American Diabetes Association. REFERENCES 'Zach, S. I., Peques, J. J., and Elliot, F. A.: Interstitial muscle capillaries in patients with diabetes mellitus: A light and electron microscope study. Metabolism 77:381-93, 1962. 2 Siperstein, M. D., Norton, W., Unger R. H., and Madison, D. L.: Diabetic capillary basement membrane width in normal, diabetic and prediabetic patients. Trans. Assoc. Am. Physicians 79:330-44, 1966. 3 Siperstein, M. D., Unger, R. H., and Madison, L. L.: Studies of muscle capillary basement membrane in normal subjects, diabetic and prediabetic patients. J. Clin. Invest. 47:1973-99, 1968. 4 Williamson, J. R., and Kilo, C : Basement membrane thickening and the mystery of diabetes. Hospital Practice: .52:109-17, 1971. 5 Williamson, J. R., Vogler, N. J., and Kilo, C : Estimation of vascular basement membrane thickness. Diabetes 78:567-78, 1969. 6 Bencosme, S. A., West, R. O., Kerr, J. W., and Willson D. L.: Diabetic capillary angiopathy in human skeletal muscle. Am. J. Med. 40:67-77, 1966. 7 Kilo, C , Vogler, N., and Williamson, J. R.: Muscle capillary basement membrane changes related to aging and diabetes. Diabetes 27:881-905, 1972. 8 Danowski, T. S., Fisher, E. R., Khurana, R. C , Nolan, S., and Stephen, T.: Muscle capillary basement membranes in juvenile diabetes mellitus. Metabolism 27:1125-32, 1972. 9 Williamson, J. R., Vogler, N. J., and Kilo, C : Microvascular diseases in diabetes. Med. Clin. N. Am. .5.5:847-60, 1971. 10 Vracko, R., and Strandness, D. E.: Basal lamina of abdominal skeletal muscle capillaries in diabetics and nondiabetics. Circulation 3.5:690-700, 1967. n Vracko, R.: Skeletal muscle capillaries in diabetics: A quantitative analysis. Circulation 47:271-83, 1970.
MAY, 1976
12
Fagerberg, S. E.: Studies in pathogenesis of diabetic neuropathy. Acta Med. Scand. 7.54:145-50, 1956. 13 Aagenaes, O., and Mae, H.: Light- and electron-microscopic study of skin capillaries of diabetics. Diabetes 70:253-59, 1961. 14 Frantizis, T. G.: The ultrastructure of capillary basement membranes in the attached gingiva of diabetic and non-diabetic patients. J. Periodontol. 42:49, 1971. 15 Bloodworth, J. M. B., Jr., Engerman, R. L., and Powers, K. L.: Experimental diabetic microangiopathy: Basement membrane statistics in the dog. Diabetes 78:455-58, 196916 Cameron, D. P., Amherdt, M., Leuenberger, P., Orci, L., and Stauffacher, W.: Microvascular alterations in chronically streptozotocin-diabetic rats. In Vascular and Neurological Changes in Early Diabetes. Camerini-Davalos, R. A., and Cole, H. S., Eds. New York, Academic Press, 1973, pp. 257-64. 17 Salazar, H., Chez, R. A., and Pardo, M.: Absence of ultrastructural changes in the basement membrane of muscle capillaries in streptozocin-induced carbohydrate intolerance in rhesus monkeys. Am. J. Pathol. 77:437-45, 1973. 18 Packer, J. T., Kraner, K. L , Rose, S. D., Stuhlman, R. A., and Nelson, L. R.: Diabetes mellitus in Mystromys albicaudatus. Arch. Pathol. 89:410-15, 1970. 19 Stuhlman, R. A., Packer, J. T., and Doyle, R. E.: Spontaneous diabetes mellitus in Mystromys albicaudatus: Repeated glucose values from 620 animals. Diabetes 27:715-21, 1972. 20 Stuhlman, R. A.: The genetic mode of transmission of spontaneous diabetes mellitus in Mystromys albicaudatus: Columbia, University of Missouri, M.S. thesis, 1971. 21 Stuhlman, R. A., Srivastava, P. K., Schmidt, G., Vorbeck, M. L., and Townsend J. F.: Characterization of diabetes mellitus in South African hamsters (Mystromys albicaudatus). Diabetologia 70:685-90, 1974. 22 Stuhlman, R. A., Packer, J. T., and Rose, S. D.: Repeated blood sampling of Mystromys albicaudatus (white tailed rat). Lab. Anim. Sci. 22:268-70, 1972. 23 Renold, A. E., and Burr, I.: The pathogenesis of diabetes mellitus: Possible usefulness of spontaneous hyperglycemic syndromes in animals. Calif. Med. 7 72:23-34, 1970. 24 Renold, A. E., and Dulin, W. E., Eds.: Spontaneous diabetes in laboratory animals. Diabetologia 3.63-286, 1967. "Williamson, J. R., Vogler, N. J., and Kilo C : Early capillary basement membrane changes in subjects with diabetes mellitus. Adv. Metab. Disord. Suppl. 2:363-67, 1973. 26 Vracko, R. and Benditt, E. P.: Manifestations of diabetes mellitus—Their possible relationship to an underlying cell defect. Am. J. Pathol. 7.5:204-22, 1974. 27 Riley, T., Stuhlman, R. A., Van Peenen, H. J., Esterly, J. A., and Townsend, J. F.: Glomerular lesions of diabetes mellitus in Mystromys albicaudatus. Arch. Pathol. 99:167-69, 1975.
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