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Failure of rostral neural tube closure in Spca1 / embryos. A, Spca1 / and Spca1 / embryos from the same litter at ED 10.5. The neural tube (white arrows) is closed in the wild type but open in the null mutant. Note the appearance of the neuroectoderm in the area of the rhombencephalon, which is ruffled in the mutant (red arrow). B, H&E-stained sections of wild-type and null mutant embryos at ED 9.5, fixed in utero. The neural tube is closed in the wild type and open in the mutant, which also exhibits ruffling of the neuroectoderm (red arrow). N, neuroectoderm; M, mesenchyme. Bar, 100 m.
Source publication
Loss of one copy of the human ATP2C1 gene, encoding SPCA1 (secretory pathway Ca(2+)-ATPase isoform 1), causes Hailey-Hailey disease, a skin disorder. We performed targeted mutagenesis of the Atp2c1 gene in mice to analyze the functions of this Golgi membrane Ca(2+) pump. Breeding of heterozygous mutants yielded a normal Mendelian ratio among embryo...
Contexts in source publication
Context 1
... Closure of Neural Tube in Null Mutant Em- bryos-The most striking gross defect in Spca1 / embryos was failure of neural tube closure (Fig. 4). During the early stages of neural tube development on ED 8.5, null mutants were morphologically indistinguishable from Spca1 / and Spca1 / embryos, and there was no apparent growth retarda- tion. Primary neurulation appeared to have proceeded nor- mally in Spca1 / embryos, with development of the notocord, neural plate, neural crests, ...
Context 2
... than in wild-type embryos, but they occurred at the appropriate interface between the mesenchyme and the open rostral neural tube. Spca1 / embryos that survived to ED 10.5 were growth-retarded and continued to have an open rostral neural tube, and ruffling of the edges of the open neural tube was observed in some of the mutants (red arrows in Fig. 4, A and B). The medial hinge point of the neural tube often appeared appropriate in Spca1 / embryos, but dorsolateral hinging was incomplete from the hind brain forward. Despite growth retardation and nonclosure of the rostral neural tube in null embryos, some growth and development continued between EDs 8.5 and 10.5, and, very ...
Similar publications
Hailey-Hailey disease (HHD) is an autosomal dominant trait characterized by erythematous and oozing skin lesions preponderantly involving the body folds. In the present unusual case, however, unilateral segmental areas along the lines of Blaschko showing a rather severe involvement were superimposed on the ordinary symmetrical phenotype. Based on t...
Citations
... Currently, no suitable HHD animal model explicitly targets ATP2C1 mutations. 13,14 Different cell models for acantholysis exhibit poor efficiency and low stability for gene downregulation. 15 A yeast-based HHD model has limitations in mimicking in vivo conditions. ...
... 21,22 Thus, we believe the intracellular Ca 2+ concentration in keratinocytes that mimic HHD requires further investigation. The embryonic lethality observed in mice with ATP2C1 mutations is probably associated with extensive cell apoptosis, 13 and Ca 2+ ions play a crucial role in mitochondrial apoptosis and cell apoptosis induction in HeLa cells. 22 Thus, we examined keratinocyte proliferation and apoptosis after ATP2C1 knockout. ...
Background
Hailey–Hailey disease (HHD) is a rare, autosomal dominant, hereditary skin disorder characterised by epidermal acantholysis. The HHD-associated gene ATPase calcium-transporting type 2C member 1 ( ATP2C1 ) encodes the protein secretory pathway Ca ²⁺ ATPase1 (SPCA1), playing a critical role in HHD pathogenesis.
Aims
We aimed to investigate the effect of ATP2C1 knockdown on keratinocytes that mimicked acantholysis in HHD.
Methods
Immunohistochemistry (IHC) was employed to evaluate the levels of cytoskeletal and tight junction proteins such as SPCA1, P-cofilin, F-actin, claudins, occludin, and zonula occludens 1 in the skin biopsies of patients with HHD. Subsequently, the expression of these proteins in cultured ATP2C1 knockdown keratinocytes was analysed using Western blotting and immunofluorescence. Furthermore, we assessed the proliferation, apoptosis, and intracellular Ca ²⁺ concentrations in the ATP2C1 -knocked keratinocytes.
Results
The results showed decreased levels of these proteins (SPCA1, P-cofilin, F-actin, claudins, occluding, and zonula occludens 1) in HHD skin lesions. Moreover, their levels decreased in human keratinocytes transfected with ATP2C1 short hairpin RNA, accompanied by morphological acantholysis. Furthermore, the proliferation and apoptosis of the keratinocytes, as well as intracellular calcium concentrations in these cells, were not affected.
Limitations
The limitations of this study are the absence of animal experiments and the failure to explore the relationship between skeletal and tight junction proteins.
Conclusion
The present study indicated that ATP2C1 inhibition led to abnormal levels of the cytoskeletal and tight junction proteins in the keratinocytes. Therefore, keratinocytes can mimic HHD-like acantholysis and serve as an in vitro model, helping develop treatment strategies against HHD.
... This mutation gives rise to cellular Ca 2+ dyshomeostasis, as SPCA1 transports Ca 2+ into the Golgi apparatus, an essential organelle for intracellular Ca 2+ storage. The disease phenotype of HHD is caused by ATP2C1 haploinsufficiency and biallelic mutations are lethal (3). Mutations in the ATP2C1 gene have been shown to produce dysfunctional proteins, that may be degraded and impair normal cellular functions, resulting in abnormal keratinocyte adhesion in the suprabasal layer of the epidermis, a feature termed acantholysis. ...
Hailey-Hailey disease is a rare hereditary skin disease caused by mutations in the ATP2C1 gene encoding the secretory pathway Ca2+/Mn2+-ATPase 1 (SPCA1) protein. Extracutaneous manifestations of Hailey-Hailey disease are plausible but still largely unknown. The aim of this study was to explore the association between Hailey-Hailey disease and diabetes. A population-based cohort study of 347 individuals with Hailey-Hailey disease was performed to assess the risks of type 1 diabetes and type 2 diabetes, using Swedish nationwide registries. Pedigrees from 2 Swedish families with Hailey-Hailey disease were also investigated: 1 with concurrent type 1 diabetes and HLA-DQ3, the other with type 2 diabetes. Lastly, a clinical cohort with 23 individuals with Hailey-Hailey disease and matched healthy controls was evaluated regarding diabetes. In the register data males with Hailey-Hailey disease had a 70% elevated risk of type 2 diabetes, whereas no excess risk among women could be confirmed. In both pedigrees an unusually high inheritance for diabetes was observed. In the clinical cohort, individuals with Hailey-Hailey disease displayed a metabolic phenotype indicative of type 2 diabetes. Hailey-Hailey disease seems to act as a synergistic risk factor for diabetes. This study indicates, for the first time, an association between Hailey-Hailey disease and diabetes and represents human evidence that SPCA1 and the Golgi apparatus may be implicated in diabetes pathophysiology.
... Currently, no suitable HHD animal model is available that speci cally targets ATP2C1 mutations [10,18]. Different cell models for acantholysis exhibit poor e ciency and low stability for gene downregulation [3]. ...
... The embryonic lethality observed in mice with ATP2C1 mutations is probably associated with extensive cell apoptosis [10], and Ca 2+ ions play a crucial role in mitochondrial apoptosis and cell apoptosis induction in HeLa cells [1]. Thus, we examined keratinocyte proliferation and apoptosis after ATP2C1 knockout. ...
Hailey–Hailey disease (HHD) is a rare, autosomal, dominant, and hereditary skin disorder characterized by epidermal acantholysis. The HHD-associated gene ATPase calcium-transporting type 2C member 1 (ATP2C1) encodes the protein secretory pathway Ca2 + ATPase1 (SPCA1), playing a critical role in HHD pathogenesis. Therefore, we aimed to investigate the effect of ATP2C1 knockdown on keratinocyte cultures that mimicked HHD. The levels of cytoskeletal and tight junction proteins such as SPCA1, P-cofilin, F-actin, claudins, occludin, and zonula occludens 1 were analyzed in skin biopsies and cultured ATP2C1 knockdown keratinocytes. The results showed decreased levels of these proteins in HHD skin lesions. Moreover, their levels decreased in human keratinocytes transfected with ATP2C1 short hairpin RNA, accompanied by morphological acantholysis. Furthermore, the proliferation and apoptosis of the keratinocytes as well as intracellular calcium concentrations in these cells were not affected. The present findings indicated that SPCA1 inhibition led to abnormal levels of the cytoskeletal and tight junction proteins in the keratinocyte cultures. Therefore, keratinocyte cultures can mimic HHD and can serve as an in vitro model, thereby helping develop treatment strategies against HHD.
... Study of these diseases is further hampered by the lack of animal models that recapitulate human disease symptoms. While mice heterozygous for Atp2a2 and Atp2c1 null alleles exist, they do not develop acantholytic skin disorders, but instead develop squamous cell carcinomas as they age (14,15). As its etiology remains unknown, no animal model exists for GD. ...
Darier, Hailey-Hailey, and Grover's diseases are rare acantholytic skin diseases. While these diseases have different underlying causes, they share defects in cell-cell adhesion in the epidermis and desmosome organization. To better understand the underlying mechanisms leading to disease in these conditions we performed RNA-seq on lesional skin samples from patients. The transcriptomic profiles of Darier, Hailey-Hailey, and Grover's disease were found to share a remarkable overlap, which did not extend to other common inflammatory skin diseases. Analysis of enriched pathways showed a shared upregulation in keratinocyte differentiation, and a decrease in cell adhesion and actin organization pathways in Darier, Hailey-Hailey, and Grover's disease. Direct comparison to atopic dermatitis and psoriasis showed that the downregulation in actin organization pathways was a unique feature in the acantholytic skin diseases. Further, upstream regulator analysis suggested that a decrease in SRF/MRTF activity was responsible for the downregulation of actin organization pathways. Staining for MRTFA in lesional skin samples showed a decrease in nuclear MRTFA in patient skin compared to normal skin. These findings highlight the significant level of similarity in the transcriptome of Darier, Hailey-Hailey, and Grover's disease, and identify decreases in actin organization pathways as a unique signature present in these conditions.
... In contrast to SERCA localized in the ER and cis-Golgi, SPCA is present in the trans-Golgi and TGN for Ca 2+ uptake, generating a large Ca 2+ gradient concentration between the ER (~500 μM) and Golgi (~100 μM) (3,7). Loss of SPCA1 is known to cause Golgi stress and apoptosis (8). In addition, loss-of-function mutations in the ATP2C1 (i.e., SPCA1) gene can cause Hailey-Hailey disease (HHD), a skin disorder characterized by persistent blisters and erosion that is inherited in an autosomal dominant manner (9). ...
Secretory pathway Ca2+/Mn2+ ATPase 1 (SPCA1) actively transports cytosolic Ca2+ and Mn2+ into the Golgi lumen, playing a crucial role in cellular calcium and manganese homeostasis. Detrimental mutations of the ATP2C1 gene encoding SPCA1 cause Hailey-Hailey disease. Here, using nanobody/megabody technologies, we determined cryo-electron microscopy structures of human SPCA1a in the ATP and Ca2+/Mn2+-bound (E1-ATP) state and the metal-free phosphorylated (E2P) state at 3.1- to 3.3-Å resolutions. The structures revealed that Ca2+ and Mn2+ share the same metal ion-binding pocket with similar but notably different coordination geometries in the transmembrane domain, corresponding to the second Ca2+-binding site in sarco/endoplasmic reticulum Ca2+-ATPase (SERCA). In the E1-ATP to E2P transition, SPCA1a undergoes similar domain rearrangements to those of SERCA. Meanwhile, SPCA1a shows larger conformational and positional flexibility of the second and sixth transmembrane helices, possibly explaining its wider metal ion specificity. These structural findings illuminate the unique mechanisms of SPCA1a-mediated Ca2+/Mn2+ transport.
... However, when given alone, anti-M3AR did not cause morphological changes in that in vitro model, while anti-SPCA1 given alone resulted in shrinkage of epidermal keratinocytes [24]. SPCA1 is located on the Golgi apparatus, and it is thought that a defect in SPCA1 Ca 2+ sequestration contributes to Golgi stress leading to apoptosis [25]. The Golgi complex is capable of transducing pro-apoptotic signals which are partially mediated through caspase 2 (Cs-2) that localizes to the Golgi apparatus [26,27]. ...
Pemphigus vulgaris (PV) is an IgG autoantibody-mediated, potentially fatal mucocutaneous disease manifested by progressive non-healing erosions and blisters. Beyond acting to inhibit adhesion molecules, PVIgGs elicit a unique process of programmed cell death and detachment of epidermal keratinocytes termed apoptolysis. Mitochondrial damage by antimitochondrial antibodies (AMA) has proven to be a critical link in this process. AMA act synergistically with other autoantibodies in the pathogenesis of PV. Importantly, absorption of AMA inhibits the ability of PVIgGs to induce blisters. Pharmacologic agents that protect mitochondrial function offer a new targeted approach to treating this severe immunoblistering disease.
... SPCA1, encoded by the ATP2C1 gene, is a Golgi-residing protein that pumps Ca 2+ from the cytosol into the Golgi lumen to be packaged into vesicles and transported in the secretory pathway (5). Mutations in the ATP2C1 gene are reported in patients with Hailey-Hailey disease, a rare skin disorder that is inherited in an autosomal dominant pattern (6). SPCA1 has also been implicated in Mn 2+ import into the Golgi; the yeast ortholog PMR1 and SPCA1 in C. elegans were shown to transport Mn 2+ with high affinity (7). ...
Manganese (II) accumulation in human brain microvascular endothelial cells (hBMVEC1) is mediated by the metal ion transporters ZIP8 and ZIP14. The plasma membrane occupancy of ZIP14, in particular, is increased in cells treated with Mn2+, lipopolysaccharide (LPS), or IL-6 but the mechanism of this regulation has not been elucidated. The calcium-transporting type 2C member 1 ATPase, SPCA1, is a Golgi-localized Ca2+-uptake transporter thought to support Golgi uptake of Mn2+ also. Here, we show using surface protein biotinylation, indirect immunofluorescence, and GFP-tagged proteins that cytoplasmic Ca2+ regulates ZIP8- and ZIP14-mediated manganese accumulation in hBMVEC by increasing the plasma membrane localization of these transporters. We demonstrate that RNAi knockdown of SPCA1 expression results in increase in cytoplasmic Ca2+ levels. In turn, we found increase in cytoplasmic Ca2+ enhances membrane-localized ZIP8 and ZIP14 and a subsequent increase in 54Mn2+ uptake. Furthermore, overexpression of WT SPCA1 or a gain-of-function mutant resulted in a decrease in cytoplasmic Ca2+ and 54Mn2+ accumulation. While addition of Ca2+ positively regulated ZIP-mediated 54Mn2+ uptake, we show chelation of Ca2+ diminished manganese transport. In conclusion, the modulation of ZIP8 and ZIP14 membrane cycling by cytoplasmic calcium is a novel finding and provides new insight into the regulation of the uptake of Mn2+ and other divalent metal ions mediated ZIP metal transporters.
... On the other hand, high Ca 2+ concentration in the TGN is required for the segregation and sorting of secretory cargo at TGN. This involves the Ca 2+ pump SPCA1 and Ca 2+ binding protein Cab45 [55,[95][96][97]. SPCA1 binds to the actin-severing protein actin-depolymerizing factor (ADF)/cofilin1 on the TGN via dynamic actin and promotes Ca 2+ influx into the TGN lumen. ...
The Golgi apparatus is a membrane organelle located in the center of the protein processing and trafficking pathway. It consists of sub-compartments with distinct biochemical compositions and functions. Main functions of the Golgi, including membrane trafficking, protein glycosylation, and sorting, require a well-maintained stable microenvironment in the sub-compartments of the Golgi, along with metal ion homeostasis. Metal ions, such as Ca2+, Mn2+, Zn2+, and Cu2+, are important cofactors of many Golgi resident glycosylation enzymes. The homeostasis of metal ions in the secretory pathway, which is required for proper function and stress response of the Golgi, is tightly regulated and maintained by transporters. Mutations in the transporters cause human diseases. Here we provide a review specifically focusing on the transporters that maintain Golgi metal ion homeostasis under physiological conditions and their alterations in diseases.
... Oxidative stress and ROS are decisive factors of UVAinduced senescence and MAPK activation. Several reports in different areas congruously found that SPCA1 was associated with oxidative stress and ROS due to its capability of regulating [Ca 2+ ]i; some refer to this as "Golgi stress" (Okunade et al., 2007;Shull et al., 2011). Keratinocytes derived from Hailey-Hailey disease patients, which lack one functional copy of the ATP2C1 gene, underwent oxidative stress, while ATP2C1 inactivation increased oxidative stress in cultured human keratinocytes (Cialfi et al., 2010(Cialfi et al., , 2016. ...
Secretory pathway calcium ATPase 1 (SPCA1) is a calcium pump localized specifically to the Golgi. Its effects on UVA-induced senescence have never been examined. In our study, expression of SPCA1 was increased in UVA-irradiated human dermal fibroblasts (HDFs) by activating mitogen-activated protein kinase (MAPK) and its downstream transcription factor, c-jun. Dual-luciferase reporter and chromatin immunoprecipitation assays revealed that c-jun regulated SPCA1 by binding to its promoter. Furthermore, downregulating SPCA1 with siRNA transfection aggravated UVA-induced senescence due to an elevation of intracellular calcium concentrations and a subsequent increase in reactive oxygen species (ROS) and MAPK activity. In contrast, overexpression of SPCA1 reduced calcium overload, consequently lowering the ROS level and suppressing MAPK activation. This alleviated the cellular senescence caused by UVA irradiation. These results indicated that SPCA1 might exert a protective effect on UVA-induced senescence in HDFs via forming a negative feedback loop. Specifically, activation of MAPK/c-jun triggered by UVA transcriptionally upregulated SPCA1. In turn, the increased SPCA1 lowered the intracellular Ca²⁺ level, probably through pumping Ca²⁺ into the Golgi, leading to a reduction of ROS, eventually decreasing MAPK activity and diminishing UVA-induced senescence.
... In neural tissue SPCA1 exhibited an important role in the control of cytoskeletal dynamics in mice neuroepithelial cells and perturbation of calcium homeostasis impaired apical constriction during neural tube closure [195]. Since the GA is an important platform for a number of signaling cascades, inactivation of SPCA1 can also induce the disturbances in mitochondrial structure and metabolism, increasing their sensitivity to stress conditions [196]. Thereby, the abnormal function of the GA in several neuropathologies can be initiated by one or more aforementioned mechanisms. ...
Calcium in mammalian neurons is essential for developmental processes, neurotransmitter release, apoptosis, and signal transduction. Incorrectly processed Ca2+ signal is well-known to trigger a cascade of events leading to altered response to variety of stimuli and persistent accumulation of pathological changes at the molecular level. To counterbalance potentially detrimental consequences of Ca2+, neurons are equipped with sophisticated mechanisms that function to keep its concentration in a tightly regulated range. Calcium pumps belonging to the P-type family of ATPases: plasma membrane Ca2+-ATPase (PMCA), sarco/endoplasmic Ca2+-ATPase (SERCA) and secretory pathway Ca2+-ATPase (SPCA) are considered efficient line of defense against abnormal Ca2+ rises. However, their role is not limited only to Ca2+ transport, as they present tissue-specific functionality and unique sensitive to the regulation by the main calcium signal decoding protein—calmodulin (CaM). Based on the available literature, in this review we analyze the contribution of these three types of Ca2+-ATPases to neuropathology, with a special emphasis on mental diseases.
