RESUMO
The analysis of neural circuits has been revolutionized by optogenetic methods. Light-gated chloride-conducting anion channelrhodopsins (ACRs)-recently emerged as powerful neuron inhibitors. For cells or sub-neuronal compartments with high intracellular chloride concentrations, however, a chloride conductance can have instead an activating effect. The recently discovered light-gated, potassium-conducting, kalium channelrhodopsins (KCRs) might serve as an alternative in these situations, with potentially broad application. As yet, KCRs have not been shown to confer potent inhibitory effects in small genetically tractable animals. Here, we evaluated the utility of KCRs to suppress behavior and inhibit neural activity in Drosophila, Caenorhabditis elegans, and zebrafish. In direct comparisons with ACR1, a KCR1 variant with enhanced plasma-membrane trafficking displayed comparable potency, but with improved properties that include reduced toxicity and superior efficacy in putative high-chloride cells. This comparative analysis of behavioral inhibition between chloride- and potassium-selective silencing tools establishes KCRs as next-generation optogenetic inhibitors for in vivo circuit analysis in behaving animals.
Assuntos
Caenorhabditis elegans , Neurônios , Optogenética , Peixe-Zebra , Animais , Caenorhabditis elegans/genética , Neurônios/metabolismo , Neurônios/fisiologia , Optogenética/métodos , Channelrhodopsins/metabolismo , Channelrhodopsins/genética , Humanos , Drosophila , Canais de Potássio/metabolismo , Canais de Potássio/genética , Cloretos/metabolismo , Animais Geneticamente Modificados , Comportamento Animal , Células HEK293 , Drosophila melanogasterRESUMO
BACKGROUND: Femoral nerve block is widely used for regional anesthesia and analgesia in many lower limb operations. Peripheral nerve stimulation of the femoral nerve may elicit 1 of 2 responses: contraction of the sartorius muscle through stimulation of its muscular branch or contraction of the quadriceps muscle through stimulation of its respective muscular branches. Historically, a quadriceps response has been preferred. We hypothesized that the success of femoral nerve block using a sartorius muscle evoked motor response is equivalent to that using quadriceps muscle twitch response. This prospective randomized double-blind controlled trial compared sartorius or quadriceps muscle evoked motor response as the end point for stimulation for femoral nerve block. METHODS: Seventy-two patients scheduled for primary unilateral total knee arthroplasty were randomly assigned to undergo femoral nerve block using either the sartorius or the quadriceps evoked muscle response as an end point of stimulation. Motor block of the femoral and sensory block of the femoral, saphenous, and lateral femoral cutaneous nerves were assessed. The primary outcome was the quality of motor and sensory block of the femoral nerve 30 minutes after injection of 20 mL of 0.5% ropivacaine. Secondary outcomes were duration of femoral nerve block, time required to perform the nerve block, total dose of hydromorphone patient-controlled analgesia, and postoperative pain by visual analog score 24 hours after block insertion. In addition, the spread of local anesthetic and the position of the needle in relation to the femoral nerve were assessed by means of ultrasonography. RESULTS: There were no statistically significant differences in the proportion of patients with either complete alone or complete and partial block combined between quadriceps and sartorius groups 30 minutes after block insertion; femoral nerve (P = 0.49; P = 0.13), the saphenous nerve (P = 0.64; P = 0.21), or the lateral femoral cutaneous nerves (P = 0.2; P = 0.35). Patient-controlled analgesia hydromorphone consumption was significantly higher in the group that underwent sartorius muscle stimulation ([mean ± SD] 4.9 ± 3.6 mg [range, 0-13.2 mg] vs 3.1 ± 2.7 mg [range, 0-10.0 mg]; P = 0.024). CONCLUSION: Our study demonstrated that using sartorius or quadriceps evoked muscle twitch as an end point of stimulation was associated with an equivalent degree of femoral nerve block.