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Odem, M. A.

Publications and source records attributed to Odem, M. A..

3 recordsLinked to original sources

The role of urothelial PIEZO ion channels in urinary function

The urothelium is increasingly recognized as a dynamic sensory interface that contributes to the regulation of urinary tract function. Mechanosensitive PIEZO ion channels expressed by urothelial cells have been proposed to play important roles in bladder fullness sensation and urinary reflexes. However, the functional contribution of urothelial PIEZO channels to these processes remains unclear. Here, we investigated the consequences of urothelial Piezo1 and Piezo2 deletion using the commonly used Upk2-cre transgenic line. Surprisingly, combined deletion of urothelial PIEZO channels did not alter urinary behavior, and mice lacking urothelial PIEZO1/2 exhibited normal bladder pressure-volume relationships, indicating preserved bladder filling and voiding function. Notably, Upk2-cre; Piezo1fl/fl mice displayed enhanced external urethral sphincter reflex activity during voiding, suggesting that loss of PIEZO1 from cells targeted by Upk2-cre may influence sensory signaling outside of the bladder urothelium. Consistent with this possibility, we identified substantial off-target Cre expression in dorsal root ganglia. These findings demonstrate that the Upk2-cre line is not restricted to the urothelium and therefore raises important considerations for interpreting associated phenotypes attributed specifically to urothelial gene deletion. Biologically, this raises the possibility that PIEZO1 does not influence bladder pressure-volume handling but might contribute to urinary reflexes.

neuroscience↗

Chronic BDNF simultaneously inhibits and unmasks superficial dorsal horn neuronal activity

Brain-derived neurotrophic factor (BDNF) is critically involved in the pathophysiology of chronic pain. However, the mechanisms of BDNF action on specific neuronal populations in the spinal superficial dorsal horn (SDH) requires further study. We used chronic BDNF treatment (200 ng/ml, 5-6 days) of defined-medium, serum-free spinal organotypic cultures to study intracellular calcium ([Ca2+]i) fluctuations. A detailed quantitative analysis of these fluctuations using the Frequency-independent biological signal identification (FIBSI) program revealed that BDNF simultaneously depressed activity in some SDH neurons while it unmasked a particular subpopulation of silent neurons causing them to become spontaneously active. Blockade of gap junctions disinhibited a subpopulation of SDH neurons and reduced BDNF-induced synchrony in BDNF-treated cultures. BDNF reduced neuronal excitability by measuring spontaneous excitatory postsynaptic currents. This was similar to the depressive effect of BDNF on the [Ca2+]i fluctuations. This study reveals novel regulatory mechanisms of SDH neuronal excitability in response to BDNF.

neuroscience↗

Frequency-independent biological signal identification (FIBSI): A free program that simplifies intensive analysis of non-stationary time series data

Extracting biological signals from non-linear, dynamic and stochastic experimental data can be challenging, especially when the signal is non-stationary. Many currently available methods make assumptions about the data structure (e.g., signal is periodic, sufficient recording time) and modify the raw data in pre-processing using filters and/or transformations. With an agnostic approach to biological data analysis as a goal, we implemented a signal detection algorithm in Python that quantifies the dimensional properties of waveform deviations from baseline via a running fit function. We call the resulting free program frequency-independent biological signal identification (FIBSI). We demonstrate the utility of FIBSI on two disparate types of experimental data: in vitro whole-cell current-clamp electrophysiological recordings of rodent sensory neurons (i.e., nociceptors) and in vivo fluorescence image time-lapse movies capturing gastrointestinal motility in larval zebrafish. In rodent nociceptors, depolarizing fluctuations in membrane potential are irregular in shape and difficult to distinguish from noise. Using FIBSI, we determined that nociceptors from naïve mice generate larger, more frequent fluctuations compared to naïve rats, suggesting species-specific specializations in rodent nociceptors. In zebrafish, measuring gut motility is a useful tool for addressing developmental and disease-related mechanisms associated with gut function. However, available methods are laborious, technically complex, and/or not cost-effective. We developed and tested a novel assay that can characterize intestinal peristalsis using imaging time series datasets. We used FIBSI to identify muscle contractions in the fluorescence signals and compared their frequencies in unfed and fed larvae. Additionally, FIBSI allowed us to discriminate between peristalsis and oscillatory sphincter-like movements in functionally distinct gut segments (foregut, midgut, and cloaca). We conclude that FIBSI, which is freely available via GitHub, is widely useful for the unbiased analysis of non-stationary signals and extraction of biologically meaningful information from experimental time series data and can be employed for both descriptive and hypothesis-driven investigations.Author Summary Biologists increasingly work with large, complex experimental datasets. Those datasets often encode biologically meaningful signals along with background noise that is recorded along with the biological data during experiments. Background noise masks the real signal but originates from other sources, for example from the equipment used to perform the measurements or environmental disturbances. When it comes to analyzing the data, distinguishing between the real biological signals and the background noise can be very challenging. Many existing programs designed to help scientists with this problem are either difficult to use, not freely available, or only appropriate to use on very specific types of datasets. The research presented here embodies our goal of helping others to analyze their data by employing a powerful but novice-friendly program that describes multiple features of biological activity in its raw form without abstract transformations. We show the program’s applicability using two different kinds of biological activity measured in our labs. It is our hope that this will help others to analyze complex datasets more easily, thoroughly, and rigorously.Competing Interest StatementThe authors have declared no competing interest.View Full Text

physiology↗