Cranial manipulation affects cholinergic pathway gene expression in aged rats

Ramu Anandakrishnan, PhD; Hope Tobey, DO; Steven Nguyen, BS; Osscar Sandoval, BS; Bradley G. Klein, PhD; and Blaise M. Costa, PhD
Notes and Affiliations
Notes and Affiliations

Received: July 14, 2021

Accepted: September 1, 2021

Published: January 10, 2022

  • Ramu Anandakrishnan, PhD, 

    Gibbs Cancer Center and Research Institute, Spartanburg, SC, USA

  • Hope Tobey, DO, 

    Sports and Osteopathic Medicine, Edward Via College of Osteopathic Medicine, Blacksburg, VA, USA

  • Steven Nguyen, BS, 

    Edward Via College of Osteopathic Medicine, Blacksburg, VA, USA

  • Osscar Sandoval, BS, 

    Edward Via College of Osteopathic Medicine, Blacksburg, VA, USA

  • Bradley G. Klein, PhD, 

    School of Neuroscience, Virginia Tech, Blacksburg, VA, USA

  • Blaise M. Costa, PhD, 

    Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Tech, Blacksburg, VA, USA

J Osteopath Med; 122(2): 95-103

Context: Age-dependent dementia is a devastating disorder afflicting a growing older population. Although pharmacological agents improve symptoms of dementia, age-related comorbidities combined with adverse effects often outweigh their clinical benefits. Therefore, nonpharmacological therapies are being investigated as an alternative. In a previous pilot study, aged rats demonstrated improved spatial memory after osteopathic cranial manipulative medicine (OCMM) treatment.

Objectives: In this continuation of the pilot study, we examine the effect of OCMM on gene expression to elicit possible explanations for the improvement in spatial memory.

Methods: OCMM was performed on six of 12 elderly rats every day for 7 days. Rats were then euthanized to obtain the brain tissue, from which RNA samples were extracted. RNA from three treated and three controls were of sufficient quality for sequencing. These samples were sequenced utilizing next-generation sequencing from Illumina NextSeq. The Cufflinks software suite was utilized to assemble transcriptomes and quantify the RNA expression level for each sample.

Results: Transcriptome analysis revealed that OCMM significantly affected the expression of 36 genes in the neuronal pathway (false discovery rate [FDR] <0.004). The top five neuronal genes with the largest-fold change were part of the cholinergic neurotransmission mechanism, which is known to affect cognitive function. In addition, 39.9% of 426 significant differentially expressed (SDE) genes (FDR<0.004) have been previously implicated in neurological disorders. Overall, changes in SDE genes combined with their role in central nervous system signaling pathways suggest a connection to previously reported OCMM-induced behavioral and biochemical changes in aged rats.

Conclusions: Results from this pilot study provide sufficient evidence to support a more extensive study with a larger sample size. Further investigation in this direction will provide a better understanding of the molecular mechanisms of OCMM and its potential in clinical applications. With clinical validation, OCMM could represent a much-needed low-risk adjunct treatment for age-related dementia including Alzheimer’s disease.

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