bioRxiv Science⌕ Search

Biology subjects

Ho, M. L.

Publications and source records attributed to Ho, M. L..

2 recordsLinked to original sources

Training-induced alterations in the modulation of human motoneuron discharge patterns with contraction force

Motoneurons adapt to both resistance and endurance training in reduced animal preparations, with adaptations seemingly more apparent in higher threshold neurons, but similar evidence in humans is lacking. Here, we compared the identified motor unit (MU) discharge patterns from decomposed electromyography signals acquired during triangular dorsiflexion contractions up to 70% of maximal voluntary force (MVF) between resistance-trained, endurance-trained, and untrained individuals (n=23 in each group). We then estimated intrinsic motoneuron properties and garnered insight about the proportion of excitatory, inhibitory, and neuromodulatory inputs contributing to motor commands across contraction intensities in each group. Participants also performed a task where a triangular contraction was superimposed onto a sustained one designed to challenge inhibitory control of dendritic persistent inward currents (PICs). Both trained groups demonstrated greater MU discharge rates with greater ascending discharge rate modulation during higher contraction forces ([≥]50% MVF), which were accompanied by more linear MU discharge patterns and greater post-acceleration attenuation slopes of the ascending discharge rates. No differences in discharge rate hysteresis or the discharge rate characteristics during the sombrero tasks between groups, suggesting no differences in neuromodulatory input. Conversely, resistance-compared to endurance-trained individuals exhibited greater acceleration slopes during lower contractions forces ([≤]50% MVF), indicating the possibility of enhanced initial activation of PICs. Collectively, the greater and more linear MU discharge patterns in the trained groups either suggests a more reciprocal (i.e., push-pull) excitation-inhibition coupling during higher contraction forces or enhanced excitatory synaptic input to the motor pool, which might underpin greater force production of trained individuals.

neuroscience↗

A pipeline for rapidly generating genetically engineered mouse models of pancreatic cancer using in vivo CRISPR-Cas9 mediated somatic recombination

Genetically engineered mouse models (GEMMs) that recapitulate the major genetic drivers in pancreatic ductal adenocarcinoma (PDAC) have provided unprecedented insights into the pathogenesis of this lethal neoplasm. Nonetheless, generating an autochthonous model is an expensive, time consuming and labor intensive process, particularly when tissue specific expression or deletion of compound alleles are involved. In addition, many of the current PDAC GEMMs cause embryonic, pancreas-wide activation or loss of driver alleles, neither of which reflects the cognate human disease scenario. The advent of CRISPR/Cas9 based gene editing can potentially circumvent many of the aforementioned shortcomings of conventional breeding schema, but ensuring the efficiency of gene editing in vivo remains a challenge. Here we have developed a pipeline for generating PDAC GEMMs of complex genotypes with high efficiency using a single \"workhorse\" mouse strain expressing Cas9 in the adult pancreas under a p48 promoter. Using adeno-associated virus (AAV) mediated delivery of multiplexed guide RNAs (sgRNAs) to the adult murine pancreas of p48-Cre; LSL-Cas9 mice, we confirm our ability to express an oncogenic Kras G12D allele through homology-directed repair (HDR), in conjunction with CRISPR-induced disruption of cooperating alleles (Trp53, Lkb1 and Arid1A). The resulting GEMMs demonstrate a spectrum of precursor lesions (pancreatic intraepithelial neoplasia [PanIN] or Intraductal papillary mucinous neoplasm [IPMN] with eventual progression to PDAC. Next generation sequencing of the resulting murine PDAC confirms HDR of oncogenic KrasG12D allele at the endogenous locus, and insertion deletion (\"indel\") and frameshift mutations of targeted tumor suppressor alleles. By using a single \"workhorse\" mouse strain and optimal AAV serotype for in vivo gene editing with combination of driver alleles, we have created a facile autochthonous platform for interrogation of the PDAC genome.

cancer biology↗