Loss-of-function mutations of progranulin are associated with frontotemporal dementia in humans, and its deficiency in mice is a model for this disease but with normal life expectancy and mild cognitive decline on aging. The present study shows that aging progranulin deficient mice develop progressive polydipsia and polyuria under standard housing conditions starting at middle age (6-9 months). They showed high water licking behavior and doubling of the normal daily drinking volume, associated with increased daily urine output and a decrease of urine osmolality, all maintained during water restriction. Creatinine clearance, urine urea, urine albumin and glucose were normal. Hence, there were no signs of osmotic diuresis or overt renal disease, other than a concentrating defect. In line, the kidney morphology and histology revealed a 50% increase of the kidney weight, kidney enlargement, mild infiltrations of the medulla with pro-inflammatory cells, widening of tubules but no overt signs of a glomerular or tubular pathology. Plasma vasopressin levels were on average about 3-fold higher than normal levels, suggesting that the water loss resulted from unresponsiveness of the collecting tubules towards vasopressin, and indeed aquaporin-2 immunofluorescence in collecting tubules was diminished, whereas renal and hypothalamic vasopressin were increased, the latter in spite of substantial astrogliosis in the hypothalamus. The data suggest that progranulin deficiency causes nephrogenic diabetes insipidus in mice during aging. Possibly, polydipsia in affected patients - eventually interpreted as psychogenic polydipsia - may point to a similar concentrating defect.
Figure 1. Body weight, drinking and feeding behavior of progranulin deficient (Grn-/-) and control mice (Grn+/+)
A) Scatter plots showing the body weight of young and aged male and female Grn-/- and Grn+/+ mice. B) Organ weights of aged Grn-/- and Grn+/+ mice. C) Number of lickings within 24h in the IntelliCage during free-drinking and restricted-drinking experiments. During ’free-drinking’, access to the water bottles was allowed for 24h on nosepoking at the doors. During ’restricted-drinking’, access to the water bottles was mostly denied except for 2x2h per day (11-12 am and 4-5 pm). D) Scatter plots showing the 24h drinking volume of aged Grn-/- and Grn+/+ mice during free drinking and water restriction for 2x12h with one-hour free drinking in between. E) Scatter plots showing the weight of food and water consumed within 24h in sex-matched young and aged Grn-/- and Grn+/+ mice in the Phenomaster cage. F) Phenomaster analysis of drinking and feeding behavior of old Grn-/- and Grn+/+ mice during presentation of tap water or sweet water with 20% sucrose (mean ± SD). The data show the food weight and drinking volume consumed within 24h. For all subpanels each scatter represents one mouse, the line is the mean and the whisker show the standard deviation (SD). Asterisks indicate statistically significant differences between genotypes (unpaired, 2-tailed Student’s t-test for each gender, organ, drinking or feeding. * P< 0.05, ** P<0.01, *** P<0.001, **** P<0.0001).
Figure 2. Renal and metabolic functions of progranulin deficient (Grn-/-) and control mice (Grn+/+)
A) Scatter plots showing the 24h urine volume, urine osmolality and urine specific gravity of aged Grn-/- and Grn+/+ mice (12-16 months old). B) Concentration of glucose in 24h-urine and plasma of aged Grn-/- and Grn+/+ mice. C) Creatinine concentrations in 24h-urine and plasma, and creatinine clearance of aged Grn-/- and Grn+/+ mice. D) Concentrations of urea and albumin in 24h-urine of aged Grn-/- and Grn+/+ mice. E) Concentrations of arginine vasopressin (AVP, antidiuretic hormone) in plasma and in crude tissue extracts of the hypothalamus of aged Grn-/- and Grn+/+ mice and immunofluorescence analysis of AVP in the kidney (bottom, scale bar 50 µm). F, G) Plasma concentrations of ghrelin and agouti related protein (Agrp) of aged Grn-/- and Grn+/+ mice. For all subpanels each scatter represents one mouse, sexes were matched between groups, the line is the mean and the whisker show the standard deviation (SD). Asterisks indicate statistically significant differences between genotypes (unpaired, 2-tailed Student’s t-test. * P< 0.05, ** P<0.01, *** P<0.001).
Figure 3. Histomorphology of the kidney of aged progranulin deficient (Grn-/-) and control mice (Grn+/+)
A) Immunostaining of myeloid-derived F48/80-positive immune cells (brown), with hematoxylin counterstaining of nuclei (blue). Immune cells were counted per field of view and averaged (10 fields per mouse of 3 mice per group). Numbers differed significantly between genotypes (unpaired, 2-tailed Student’s t-test). Scale bars 50 µm. B) Periodic acid-Schiff (PAS) staining of polysaccharides and mucous substances. Histomorphometric scores did not differ between genotypes, except for a higher number of immune cells in Grn-/- mice. Scale bars 50 µm. C, D) Concentrations of C-reactive protein (CRP) and zinc in plasma of aged Grn-/- and Grn+/+ mice. Asterisks indicate statistically significant differences between genotypes (unpaired, 2-tailed Student’s t-test, **** P<0.0001).
Figure 4. Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, AQP2, AQP4) in the kidney of aged progranulin deficient (Grn-/-) and control mice (Grn+/+)
A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P<0.05, ***P<0.001).
Figure 5. Immunofluorescence analysis of arginine vasopressin (AVP), glial fibrillary acidic protein (GFAP) of astrocytes and DAPI counterstain of nuclei in the hypothalamus of aged progranulin deficient (Grn-/-) and control mice (Grn+/+)
The Gensat images in the right give an overview of the localization of the paraventricular nucleus (PVN) and the nucleus supraopticus (SON), which are the major sites for vasopressin producing neurons. The upper panels show AVP, GFAP and DAPI in the PVN as overview and zoom-in images, and the lower panels show the SON. AVP immunofluorescence was more intense in Grn-/- and AVP positive neurons appear to be enlarged. GFAP staining reveals astrogliosis in Grn-/-. Scale bars 300 µm for overviews and 100 µm for zoom-in images.
Table 1 Serum and urine electrolytes in progranulin knockout (Grn-/-) and wildtype (Grn+/+) female mice.
Cruts M, Gijselinck I, van der Zee J, Engelborghs S, Wils H, Pirici D, Rademakers R, et al. (2006). Null mutations in progranulin cause ubiquitin-positive frontotemporal dementia linked to chromosome 17q21. Nature, 442: 920-4.
Baker M, Mackenzie IR, Pickering-Brown SM, Gass J, Rademakers R, Lindholm C, et al. (2006). Mutations in progranulin cause tau-negative frontotemporal dementia linked to chromosome 17. Nature, 442: 916-9.
Gotzl JK, Mori K, Damme M, Fellerer K, Tahirovic S, Kleinberger G, et al. (2014). Common pathobiochemical hallmarks of progranulin-associated frontotemporal lobar degeneration and neuronal ceroid lipofuscinosis. Acta Neuropathol, 127: 845-60.
Mackenzie IR, Baker M, Pickering-Brown S, Hsiung GY, Lindholm C, Dwosh E, Gass J, Cannon A, Rademakers R, Hutton M, Feldman HH (2006). The neuropathology of frontotemporal lobar degeneration caused by mutations in the progranulin gene. Brain, 129: 3081-90.
Hardt S, Heidler J, Albuquerque B, Valek L, Altmann C, Wilken-Schmitz A, Schafer MKE, Wittig I, Tegeder I (2017). Loss of synaptic zinc transport in progranulin deficient mice may contribute to progranulin-associated psychopathology and chronic pain. Biochim Biophys Acta. 1863:2727-2745
Yin F, Dumont M, Banerjee R, Ma Y, Li H, Lin MT, Beal MF, Nathan C, Thomas B, Ding A (2010). Behavioral deficits and progressive neuropathology in progranulin-deficient mice: a mouse model of frontotemporal dementia. FASEB J, 24: 4639-47.
Ahmed Z, Sheng H, Xu YF, Lin WL, Innes AE, Gass J, et al. (2010). Accelerated lipofuscinosis and ubiquitination in granulin knockout mice suggest a role for progranulin in successful aging. Am J Pathol, 177: 311-24.
Albuquerque B, Haussler A, Vannoni E, Wolfer DP, Tegeder I (2013). Learning and memory with neuropathic pain: impact of old age and progranulin deficiency. Front Behav Neurosci, 7: 174.
Filiano AJ, Martens LH, Young AH, Warmus BA, Zhou P, Diaz-Ramirez G, Jiao J, Zhang Z, Huang EJ, Gao FB, Farese RVJr., Roberson ED (2013). Dissociation of frontotemporal dementia-related deficits and neuroinflammation in progranulin haploinsufficient mice. J Neurosci, 33: 5352-61.
Hu F, Padukkavidana T, Vaegter CB, Brady OA, Zheng Y, Mackenzie IR, Feldman HH, Nykjaer A, Strittmatter SM (2010). Sortilin-mediated endocytosis determines levels of the frontotemporal dementia protein, progranulin. Neuron, 68: 654-67.
He Z, Bateman A (1999). Progranulin gene expression regulates epithelial cell growth and promotes tumor growth in vivo. Cancer Res, 59: 3222-9.
Altmann C, Vasic V, Hardt S, Heidler J, Haussler A, Wittig I, Schmidt MH, Tegeder I (2016). Progranulin promotes peripheral nerve regeneration and reinnervation: role of notch signaling. Mol neurodegener, 11: 69.
Neill T, Buraschi S, Goyal A, Sharpe C, Natkanski E, Schaefer L, Morrione A, Iozzo RV (2016). EphA2 is a functional receptor for the growth factor progranulin. J Cell Biol, 215: 687-703.
Gass J, Lee WC, Cook C, Finch N, Stetler C, Jansen-West K, Lewis J, Link CD, Rademakers R, Nykjaer A, Petrucelli L (2012). Progranulin regulates neuronal outgrowth independent of Sortilin. Mol Neurodegener, 7: 33.
Sikora J, Harzer K, Elleder M (2007). Neurolysosomal pathology in human prosaposin deficiency suggests essential neurotrophic function of prosaposin. Acta Neuropathol, 113: 163-75.
Zhou X, Sun L, Bastos de Oliveira F, Qi X, Brown WJ, Smolka MB, Sun Y, Hu F (2015). Prosaposin facilitates sortilin-independent lysosomal trafficking of progranulin. J Cell Biol, 210: 991-1002.
Matsuda J, Kido M, Tadano-Aritomi K, Ishizuka I, Tominaga K, Toida K, Takeda E, Suzuki K, Kuroda Y (2004). Mutation in saposin D domain of sphingolipid activator protein gene causes urinary system defects and cerebellar Purkinje cell degeneration with accumulation of hydroxy fatty acid-containing ceramide in mouse. Hum Mol Genet, 13: 2709-23.
Sun Y, Witte DP, Zamzow M, Ran H, Quinn B, Matsuda J, Grabowski GA (2007). Combined saposin C and D deficiencies in mice lead to a neuronopathic phenotype, glucosylceramide and alpha-hydroxy ceramide accumulation, and altered prosaposin trafficking. Hum Mol Genet, 16: 957-71.
Matsuda J, Yoneshige A, Suzuki K (2007). The function of sphingolipids in the nervous system: lessons learnt from mouse models of specific sphingolipid activator protein deficiencies. J Neurochem, 103 Suppl 1: 32-8.
He Z, Ong CH, Halper J, Bateman A (2003). Progranulin is a mediator of the wound response. Nat Med, 9: 225-9.
Tang W, Lu Y, Tian QY, Zhang Y, Guo FJ, Liu GY, et al. (2011). The growth factor progranulin binds to TNF receptors and is therapeutic against inflammatory arthritis in mice. Science, 332: 478-84.
Zhou M, Tang W, Fu Y, Xu X, Wang Z, Lu Y, Liu F, Yang X, Wei X, Zhang Y, Liu J, Geng X, Zhang C, Wan Q, Li N, Yi F (2015). Progranulin protects against renal ischemia/reperfusion injury in mice Kidney Int, 87: 918-29.
Altmann C, Hardt S, Fischer C, Heidler J, Lim HY, Haussler A, et al. (2016). Progranulin overexpression in sensory neurons attenuates neuropathic pain in mice: Role of autophagy. Neurobiol Dis, 96: 294-311.
Tanaka Y, Chambers JK, Matsuwaki T, Yamanouchi K, Nishihara M (2014). Possible involvement of lysosomal dysfunction in pathological changes of the brain in aged progranulin-deficient mice. Acta neuropathol commun, 2: 78.
Perez Y, Shorer Z, Liani-Leibson K, Chabosseau P, Kadir R, Volodarsky M, Halperin D, Barber-Zucker S, Shalev H, Schreiber R, Gradstein L, Gurevich E, Zarivach R, Rutter GA, Landau D, Birk OS (2017). SLC30A9 mutation affecting intracellular zinc homeostasis causes a novel cerebro-renal syndrome. Brain, 140: 928-939.
Hisaki H, Matsuda J, Tadano-Aritomi K, Uchida S, Okinaga H, Miyagawa M, Tamamori-Adachi M, Iizuka M, Okazaki T (2012). Primary polydipsia, but not accumulated ceramide, causes lethal renal damage in saposin D-deficient mice. Am J Physiol Renal Physiol, 303: F1049-59.
Kim HK, Shin MS, Youn BS, Namkoong C, Gil SY, Kang GM, Yu JH, Kim MS (2011). Involvement of progranulin in hypothalamic glucose sensing and feeding regulation. Endocrinology, 152: 4672-82.
Suzuki M, Nishiahara M (2002). Granulin precursor gene: a sex steroid-inducible gene involved in sexual differentiation of the rat brain. Mol Genet Metab, 75: 31-7.
Zhou B, Li H, Liu J, Xu L, Guo Q, Sun H, Wu S (2015). Progranulin induces adipose insulin resistance and autophagic imbalance via TNFR1 in mice. J Mol Endocrinol, 55: 231-43.
Nguyen AD, Nguyen TA, Martens LH, Mitic LL, Farese RVJr., (2013). Progranulin: at the interface of neurodegenerative and metabolic diseases. Trends Endocrinol Metab, 24: 597-606.
Tegeder I (2016). Yeast-2-Hybrid data file showing progranulin interactions in human fetal brain and bone marrow libraries. Data Brief, 9: 1060-1062.
Norregaard R, Tao S, Nilsson L, Woodgett JR, Kakade V, Yu AS, Howard C, Rao R (2015). Glycogen synthase kinase 3alpha regulates urine concentrating mechanism in mice. Am J Physiol Renal Physiol, 308: F650-60.
Yin F, Banerjee R, Thomas B, Zhou P, Qian L, Jia T, Ma X, Ma Y, Iadecola C, Beal MF, Nathan C, Ding A (2010). Exaggerated inflammation, impaired host defense, and neuropathology in progranulin-deficient mice. J Exp Med, 207: 117-28.
Voikar V, Colacicco G, Gruber O, Vannoni E, Lipp HP, Wolfer DP (2010). Conditioned response suppression in the IntelliCage: assessment of mouse strain differences and effects of hippocampal and striatal lesions on acquisition and retention of memory. Behav Brain Res, 213: 304-12.
Krackow S, Vannoni E, Codita A, Mohammed AH, Cirulli F, Branchi I, Alleva E, Reichelt A, Willuweit A, Voikar V, Colacicco G, Wolfer DP, Buschmann JU, Safi K, Lipp HP (2010). Consistent behavioral phenotype differences between inbred mouse strains in the IntelliCage. Genes Brain Behav, 9: 722-31.
Moreth K, Frey H, Hubo M, Zeng-Brouwers J, Nastase MV, Hsieh LT, Haceni R, Pfeilschifter J, Iozzo RV, Schaefer L (2014). Biglycan-triggered TLR-2- and TLR-4-signaling exacerbates the pathophysiology of ischemic acute kidney injury. Matrix Biology, 35: 143-51.
Naccarato R, Rizzo A, Sirigu F, Bertaglia E, Previato G, Fiaschi E (1976). Renal histologic and ultrastructural findings in psychogenic polydipsia and diabetes insipidus. Nephron, 16: 226-35.
Schliebe N, Strotmann R, Busse K, Mitschke D, Biebermann H, Schomburg L, et al. (2008). V2 vasopressin receptor deficiency causes changes in expression and function of renal and hypothalamic components involved in electrolyte and water homeostasis. Am J Physiol Renal Physiol, 295: F1177-90.
Sohara E, Rai T, Yang SS, Uchida K, Nitta K, Horita S, Ohno M, Harada A, Sasaki S, Uchida S (2006). Pathogenesis and treatment of autosomal-dominant nephrogenic diabetes insipidus caused by an aquaporin 2 mutation. Proc Natl Acad Sci U S A, 103: 14217-22.
Matsuwaki T, Asakura R, Suzuki M, Yamanouchi K, Nishihara M (2011). Age-dependent changes in progranulin expression in the mouse brain. J Reprod Dev, 57: 113-9.
Shi G, Somlo D, Kim GH, Prescianotto-Baschong C, Sun S, Beuret N, Long Q, Rutishauser J, Arvan P, Spiess M, Qi L (2017). ER-associated degradation is required for vasopressin prohormone processing and systemic water homeostasis. J Clin Invest. 127(10): 3897-3912
Yang B, Zhao D, Qian L, Verkman AS (2006). Mouse model of inducible nephrogenic diabetes insipidus produced by floxed aquaporin-2 gene deletion. Am J Physiol Renal Physiol, 291: F465-72.
Poulsen SB, Kristensen TB, Brooks HL, Kohan DE, Rieg T, Fenton RA (2017). Role of adenylyl cyclase 6 in the development of lithium-induced nephrogenic diabetes insipidus. JCI insight, 2: e91042.
Cano-Penalver JL, Griera M, Serrano I, Rodriguez-Puyol D, Dedhar S, de Frutos S, Rodriguez-Puyol M (2014). Integrin-linked kinase regulates tubular aquaporin-2 content and intracellular location: a link between the extracellular matrix and water reabsorption. FASEB J, 28: 3645-59.
Hozawa S, Holtzman EJ, Ausiello DA (1996). cAMP motifs regulating transcription in the aquaporin 2 gene Am J Physiol, 270: C1695-702.
Birnbaumer M, Gilbert S, Rosenthal W (1994). An extracellular congenital nephrogenic diabetes insipidus mutation of the vasopressin receptor reduces cell surface expression, affinity for ligand, and coupling to the Gs/adenylyl cyclase system. Mol Endocrinol, 8: 886-94.
Valtin H, Coffey AK, O’Sullivan DJ, Homma S, Dousa TP (1990). Causes of the urinary concentrating defect in mice with nephrogenic diabetes insipidus. Physiologia Bohemoslovaca, 39: 103-11.
Rao R, Patel S, Hao C, Woodgett J, Harris R (2010). GSK3beta mediates renal response to vasopressin by modulating adenylate cyclase activity. J Am Soc Nephrol, 21: 428-37.
Kortenoeven ML, Pedersen NB, Miller RL, Rojek A, Fenton RA (2013). Genetic ablation of aquaporin-2 in the mouse connecting tubules results in defective renal water handling. J Physiol, 591: 2205-19.
Ho HT, Chung SK, Law JW, Ko BC, Tam SC, Brooks HL, Knepper MA, Chung SS (2000). Aldose reductase-deficient mice develop nephrogenic diabetes insipidus. Mol Cell Biol, 20: 5840-6.
Tsumura K, Li X, Murdiastuti K, Parvin MN, Akamatsu T, Yao C, Kanamori N, Inenaga K, Yamashita H, Hosoi K (2006). Downregulation of AQP2 expression in the kidney of polydipsic STR/N mice. Am J Physiol Renal Physiol, 290: F478-85.
Kutscher CL, Miller M, Schmalbach NL (1975). Renal deficiency associated with diabetes insipidus in the SWR/J mouse. Physiol Behav, 14: 815-8.
Preisser L, Teillet L, Aliotti S, Gobin R, Berthonaud V, Chevalier J, Corman B, Verbavatz JM (2000). Downregulation of aquaporin-2 and -3 in aging kidney is independent of V(2) vasopressin receptor. Am J Physiol Renal Physiol, 279: F144-52.
Hoque M, Young TM, Lee CG, Serrero G, Mathews MB, Pe’ery T (2003). The growth factor granulin interacts with cyclin T1 and modulates P-TEFb-dependent transcription. Mol Cell Biol, 23: 1688-702.
Chen-Plotkin AS, Geser F, Plotkin JB, Clark CM, Kwong LK, Yuan W, Grossman M, Van Deerlin VM, Trojanowski JQ, Lee VM (2008). Variations in the progranulin gene affect global gene expression in frontotemporal lobar degeneration. Hum Mol Genet, 17: 1349-62.
Bateman A, Bennett HP (1998). Granulins: the structure and function of an emerging family of growth factors. J Endocrinol, 158: 145-51.
Jeong HW, Jeon US, Koo BK, Kim WY, Im SK, Shin J, Cho Y, Kim J, Kong YY (2009). Inactivation of Notch signaling in the renal collecting duct causes nephrogenic diabetes insipidus in mice. J Clin Invest, 119: 3290-300.
Gooch JL, Guler RL, Barnes JL, Toro JJ (2006). Loss of calcineurin Aalpha results in altered trafficking of AQP2 and in nephrogenic diabetes insipidus. J Cell Sci, 119: 2468-76.
Kotliarova S, Jana NR, Sakamoto N, Kurosawa M, Miyazaki H, Nekooki M, Doi H, Machida Y, Wong HK, Suzuki T, Uchikawa C, Kotliarov Y, Uchida K, Nagao Y, Nagaoka U, Tamaoka A, Oyanagi K, Oyama F, Nukina N (2005). Decreased expression of hypothalamic neuropeptides in Huntington disease transgenic mice with expanded polyglutamine-EGFP fluorescent aggregates. J Neurochem, 93: 641-53.
Wood NI, Goodman AO, van der Burg JM, Gazeau V, Brundin P, Bjorkqvist M, Petersen A, Tabrizi SJ, Barker RA, Morton AJ (2008). Increased thirst and drinking in Huntington’s disease and the R6/2 mouse. Brain Res Bull, 76: 70-9.
Tanaka Y, Suzuki G, Matsuwaki T, Hosokawa M, Serrano G, Beach TG, Yamanouchi K, Hasegawa M, Nishihara M (2017). Progranulin regulates lysosomal function and biogenesis through acidification of lysosomes. Hum Mol Genet, 26(5):969-988