When an equal volume (4.5 L) of the protein solutions was used, more than three-fold excess of CA2 protein (22.5 ng) over CA1 protein (6.42 ng) was observed around the gel (Supplementary Materials, Physique S1A, indicated by the intensity of SYPRO Ruby-stained band between Lane 1 and Lane 4). during the neonatal period and early childhood [35]. CA6 is usually initially described as a gustatory protein and highly expressed in the salivary and mammary glands. Mice deficient in both copies of the gene prefer bitter taste [36,37]. Polymorphism in the human gene was also linked to bitter taste perception [37]. CARPs are predominantly expressed in neural tissues [38]. Patients with mutations in show phenotype in cerebellar ataxia, mental retardation, and AZ82 disequilibrium syndrome [19], while mice exhibit motor dysfunction and altered calcium dynamics in cerebellar granule cells [39]. Inactivation of in zebrafish leads to abnormal embryonic development and altered movement pattern [40]. CA1 is usually a very early marker for erythroid differentiation and the second most abundant non-heme protein in erythrocytes [41]. Its expression is also detected in intestinal, vascular and corneal epithelia, synovium, and cardiac capillary endothelial cells [42,43,44,45,46]. CA1-immunoreactivity was observed in both Type I and II cells in the rat carotid body [47]. Only in one study, CA1 mRNA level in the mouse brain was reported to be extremely low compared with that of CA2 [48]. Therefore, whether CA1 is usually expressed in the CNS is usually unclear. To date, there have been a total of seven studies reporting CA1 being the key protein as AZ82 a result of unbiased screenings between normal and pathological conditions [42,43,46,49,50,51,52]. The elevated CA1 level was found in the vitreous of diabetic retinopathy which contributes to retinal hemorrhage and erythrocyte lysis via prekallikrein activation [43,53]. The increased expression of CA1 found in the synovium of patients with ankylosing spondylitis may promote dysregulated calcification and bone resorption [42]. CA1 was found to be the major antigen in cecal bacterial Ag, which is usually associated with inflammatory bowel disease. The dendritic cell-mediated CA1-specific production of regulatory T cells can suppress the development of colitis induced by CD4+CD25? T cells [52]. CA1, together with CA2, are increased in diabetic ischemic cardiomyopathy, and CA1 can affect apoptosis in vitro [46]. Similar to CA2, CA1 has been shown to be a potential novel biomarker for early stage of non-small cell lung cancer [54]. In the current study, we report CA1 expression in spinal cord motor neurons. In addition, a proportion of CA1s are associated with subcellular endoplasmic reticular (ER) structures. CA1 protein levels were preferentially increased in the spinal cord of patients with amyotrophic lateral sclerosis (ALS), while CA2 did not change in these same patients. Our in vitro cell culture data exhibited that intracellularly expressed CA1 can induce apoptosis. Our study establishes CA1 expression in the human spinal cord and suggests that CA1 may have an important function in motor neuron degeneration in ALS. 2. Results 2.1. Carbonic Anhydrase I (CA1) Is usually Expressed in Human Spinal Cord Motor Neurons Since CA1 has not been reported to be expressed in the AZ82 CNS and we are interested in the potential function of CA1 in motor neurons as well as in motor neuron degeneration in the context of ALS-related Rabbit Polyclonal to SLC25A12 pathology, we first examined whether CA1 is usually expressed in human spinal cord motor neurons. Because CA2 is known to be the most abundant CA isoform in the CNS and human CA1 (hCA1) shares 59.8% identity in the amino acid sequence with human CA2 (hCA2), we would like to be certain that this CA1 antibodies used in this study were CA1-specific and did not cross-react with CA2. For this purpose, commercially available recombinant human CA1 (rhCA1) and CA2 (rhCA2) proteins were used in the Western blot analysis (Supplementary Materials, Physique S1). When an equal volume (4.5 L) of the protein solutions was used, more than three-fold excess of CA2 protein (22.5 ng) over CA1 protein (6.42 ng) was observed around the gel (Supplementary Materials, Physique S1A, indicated by the intensity of SYPRO Ruby-stained band between Lane 1 and Lane 4). Four identical blots with an equal volume (9.5 L) of CA1 and CA2 samples loaded for each lane were then probed with three different sources of the commercially available CA1 antibodies as well as a CA2 antibody. All CA1 antibodies recognized rhCA1 without any detectable cross-immunoreactivity to rhCA2 while the CA2 antibody recognized rhCA2 only (Supplementary Materials, Physique S1B). The AZ82 specificity of one of.