Merck

Bone status of acetylcholinesterase-knockout mice.

International immunopharmacology (2015-08-08)
Vivien Kauschke, Mathias Kneffel, Wolfgang Floel, Sonja Hartmann, Marian Kampschulte, Lutz Dürselen, Anita Ignatius, Reinhard Schnettler, Christian Heiss, Katrin Susanne Lips
ABSTRACT

Acetylcholinesterase (AChE) hydrolyzes acetylcholine (ACh) to acetate and choline and thereby terminates nerve impulse transmission. ACh is also expressed in bone tissue and enhances here proliferation and differentiation of osteoblasts, which makes it interesting to investigate effects of AChE deficiency on bone. To our knowledge, this is the first study that analyzed bone of heterozygous acetylcholinesterase-knockout (AChE-KO) mice. Tibia, femur, thoracic and lumbar vertebrae of 16-week-old female heterozygous AChE-KO mice and their corresponding wildtypes (WT) were analyzed using real-time RT-PCR, dual-energy X-ray absorptiometry, biomechanics, micro-computed tomography, histology and histomorphometry. Our data revealed that heterozygous AChE-KO did not cause negative effects upon bone parameters analyzed. In contrast, the number of osteoclasts per perimeter was significantly reduced in lumbar vertebrae. In addition, we found a significant decrease in trabecular perimeter of lumbar vertebrae and cortical area fraction (Ct.Ar/Tt.Ar) in the mid-diaphysis of femurs of AChE-KO mice compared to their WT. Therefore, presumably a local homozygous knockout of AChE or AChE-inhibitor administration might be beneficial for bone formation due to ACh accumulation. However, many other bone parameters analyzed did not differ statistically significantly between AChE-KO and WT mice. That might be reasoned by the compensating effect of butyrylcholinesterase (BChE).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Glutaraldehyde solution, 50 wt. % in H2O
Sigma-Aldrich
Methyl Green, zinc chloride salt, <0.5% crystal violet, Dye content 80 %
Sigma-Aldrich
Methyl Green, zinc chloride salt, certified by the Biological Stain Commission, Dye content 85 %
Sigma-Aldrich
Aluminum sulfate, 99.99% trace metals basis
Sigma-Aldrich
Sodium carbonate, anhydrous, powder, 99.999% trace metals basis
Sigma-Aldrich
Glutaric dialdehyde solution, 50 wt. % in H2O, FCC
Sigma-Aldrich
Sodium carbonate, BioUltra, anhydrous, ≥99.5% (calc. on dry substance, T)
Sigma-Aldrich
Glycerol solution, 83.5-89.5% (T)
Sigma-Aldrich
Methyl Green, zinc chloride salt, for microscopy (Bact., Bot., Hist.)
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 50% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 8% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
Sigma-Aldrich
Nitrotetrazolium Blue chloride, ≥90.0% (HPLC)
Sigma-Aldrich
Glycerin, meets USP testing specifications
Sigma-Aldrich
Nitrotetrazolium Blue chloride, powder, electrophoresis grade
Sigma-Aldrich
Sodium carbonate, BioXtra, ≥99.0%
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 70% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 25% in H2O, specially purified for use as an electron microscopy fixative
Sigma-Aldrich
Glutaraldehyde solution, Grade II, 25% in H2O
Sigma-Aldrich
Glutaraldehyde solution, 50% in H2O, suitable for photographic applications
Sigma-Aldrich
Ser-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-Lys-Tyr-Glu-Pro-Phe, ≥97% (HPLC)
Sigma-Aldrich
Methyl Green, zinc chloride salt, ~85%
Sigma-Aldrich
Sodium carbonate, anhydrous, powder or granules, free-flowing, Redi-Dri, ACS reagent, ≥99.5%
Sigma-Aldrich
Sodium carbonate, anhydrous, free-flowing, Redi-Dri, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Sodium carbonate, anhydrous, free-flowing, Redi-Dri, ACS reagent (primary standard), 99.95-100.05% dry basis
Sigma-Aldrich
Sodium carbonate, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Glycerol solution, puriss., meets analytical specification of Ph. Eur., BP, 84-88%
Sigma-Aldrich
Sodium carbonate, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99.5%
Sigma-Aldrich
Sodium carbonate, puriss., meets analytical specification of Ph. Eur., BP, NF, FCC, E500, anhydrous, 99.5-100.5% (calc. to the dried substance)
Sigma-Aldrich
Sodium carbonate, ACS reagent, anhydrous, ≥99.5%, powder or granules
Sigma-Aldrich
Sodium carbonate, ACS reagent (primary standard), anhydrous, 99.95-100.05% dry basis