Characterization of human reflex tear proteome reveals high expression of lacrimal proline‐rich protein 4 (PRR4)

In‐depth studies on the proteome of reflex tears are still inadequate. Hence, further studies on this subject will unravel the key proteins which are conjectured to possess vital functions in the protection of the ocular surface. Therefore, this study investigated the differences in the expression levels in proteome of reflex compared to basal tears. Basal (n = 10) and reflex (n = 10) tear samples from healthy subjects were collected employing the capillary method, subsequently pooled and the proteomes were characterized employing 1DE combined with LC‐ESI‐MS/MS strategy for label‐free quantitative (LFQ) analysis. The differentially expressed proteins were validated by 2DE combined with LC‐ESI‐MS/MS and targeted‐MS approach called accurate inclusion mass screening (AIMS) strategies. The analysis of the reflex tear proteome demonstrated increased abundance in proline‐rich protein 4 (PRR4) and zymogen granule protein 16 homolog B (ZG16B) for the first time. Other abundant lacrimal proteins, e.g. lactotransferrin and lysozyme remained constant. Predominantly, the lacrimal gland‐specific PRR4 represents the major increased protein in reflex tears in an attempt to wash out irritants that come into contact with the eye. Conversely, decreased abundance in Ig alpha‐1 chain C, polymeric immunoglobulin receptor, cystatin S/SN, clusterin and mammaglobin were observed. This study had further unraveled the intricate proteome regulation during reflex tearing, especially the potential role of PRR4, which may be the key player in the protection and maintenance of dynamic balance of the ocular surface.     

Structural adaptations and mechanism of reflex bleeding in the larvae of the myrmecophilous ladybird Diomus thoracicus

Reflex bleeding is an effective defensive mechanism against predators. When attacked, some insects emit hemolymph, which coagulates, quickly entangling their aggressor. Bleeding occurs at weak intersegmental membranes or through dedicated organs, which can be associated or not with glandular cells.Here, we describe the behavior and morphological structures involved in reflex bleeding in the larvae of the ladybird, Diomus thoracicus, which are intranidal parasites of the ant Wasmannia auropunctata. The larvae are tolerated by the ants thanks to odor mimicry, but some rare aggressive ant behaviors were observed that trigger reflex bleeding both at a pair of thoracic tubercles and a pair of posterodorsal abdominal humps. No glandular structure was found in association with these emission points, which suggests that the material emitted was hemolymph only. A 3D reconstruction suggested that reflex bleeding seems to be controlled by muscles whose contraction increases the internal hydrostatic pressure and pushes the hemolymph into a funnel-like structure with an opening to the outside. In D. thoracicus, the morphological structures involved in reflex bleeding are among the most complex and prominent described to date.     

The proenkephalin A fragment metorphamide shows supraspinal and spinal opioid activity in vivo

Metorphamide (Tyr-Gly-Gly-Phe-Met-Arg-Arg-Val-NH2) a novel amidated octapeptide fragment of proenkephalin A was synthesized, purified and subsequently shown to inhibit the reflex contractions of the rat urinary bladder following intracerebroventricular and spinal intrathecal microinjections. The effects of metorphamide were consistently antagonized by naloxone but not by the delta-opioid receptor antagonist ICI 174,864. Comparison of metorphamide with other proenkephalin A fragments suggested that the activity of this peptide was not due to in vivo processing to other active fragments. These data suggest that metorphamide has potent in vivo mu-opioid activity but little delta-opioid receptor activity.     

Maternal care of rabbits in the lab and on the farm: endocrine regulation of behavior and productivity

Maternal behavior in rabbits has been well described in the wild, the laboratory, and the farm. Salient characteristics include: (a) the construction of a nest (inside an underground burrow or a box), composed of straw/grass and body hair and (b) the display of a single, brief (ca. 3 min) nursing bout per day. The onset and decline of nest-building in mid and late pregnancy are controlled by specific combinations of estradiol, progesterone, testosterone, and prolactin. Following parturition (kindling) does can mate and become pregnant again. Management strategies have been used on the farm to enhance productivity, impinging on specific reproductive processes (e.g., use of hormones to synchronize estrus, artificial insemination at kindling, doe-litter separation on specific days of lactation to increase sexual receptivity and fertility). Knowledge about the rabbit's reproductive physiology and behavior will be enriched by integrating research coming from the laboratory (where estrus, pregnant-only, or lactating-only animals are the main categories investigated) with studies performed on the farm or research station (where pregnant-lactating rabbits are prevalent). Similarly, the high productivity demands of modern rabbit farms will benefit from the information obtained in the laboratory where specific issues in reproductive neuroendocrinology are explored with methodologies that are not amenable for farm use.     

Variability of motoneuron activation and the modulation of force production in a postural reflex of the hermit crab abdomen

The tri-phasic reflex in hermit crab (Pagurus pollicarus) abdomen is triggered by local mechanoreceptors and is essential for postural control. The reflex consists of three stereotypical phases: a brief, high-frequency burst, a transient cessation of firing, and a late-discharge that is much lower in frequency than the initial burst. To better understand the reflex generation of force, variability of motoneuron discharge in each of five parameters of reflex activation was assessed. An intracellular current injection routine was used to correlate each of these parameters with force production. Phase 3 motoneuron firing frequency showed the greatest correlation with force production. Phase 3 spike rate increased as a function of phase 2 duration, but the relationship between phase 2 duration and force produced by the reflex was weak. Junction potential amplitude decreased as phase 2 duration increased, and we hypothesize that this trend counteracts the increased phase 3 frequency, explaining the weak relationship of phase 2 duration and force production. Surprisingly, when phase 3 frequency was held constant and phase 2 was increased in duration, the concurrent decrease in junction potential amplitude did not reduce force production.     

Avoidance reflexes mediated by contact chemoreceptors on the legs of locusts

Taste receptors, or basiconic sensilla, are distributed over the legs of the locust and respond to direct contact with chemical stimulants. The same chemosensory neurones that responded to contact with salt solutions also responded to particular acidic odours. Odours of food and other chemicals had no effect on the chemosensory neurones. In locusts free to move, an acid odour presented to the tarsus of a hind leg evoked a rapid avoidance movement in which the tarsus was levated, the tibia flexed and the femur levated. Intracellular recordings from motor neurones that innervate muscles of the hind leg showed that when an acid odour was directed towards basiconic sensilla on the leg there was a reciprocal activation of antagonistic motor pools that move the leg segments about each joint. Thus an extensor tibiae motor neurone was inhibited while a flexor tibiae motor neurone was excited, and the tarsal depressor and retractor unguis motor neurones were inhibited while the tarsal levator motor neurone was excited. This method of odour stimulation of taste receptors generates less adaptation than direct contact with chemicals, and therefore represents an ideal method for stimulating taste receptors for further studies on the central pathways processing taste signals. Accepted: 2 June 1998     

Effects of a larval antipredator response and larval diet on adult phenotype in an aposematic ladybird beetle

Many ladybird beetles respond to a potential predation event by `reflex bleeding' or secreting a noxious defensive chemical that is similar to hemolymph. Both adults and larvae show this response. Reflex bleeding is known to reduce predator attack rates and increase prey survival after an attack, especially when reflex bleeding is employed in combination with other cues such as odor and warning coloration. In this experiment, we examined how variability in the number of reflex bleeding events and food quality in the larval stage of the aposematic ladybird beetle Harmonia axyridis affected elytral color, development time, and terminal size in adults. Effects of reflex bleeding were subtle and may have been influenced by diet treatments. Adult color did not differ between bleed treatment groups but beetles that reflex bled tended to take longer to develop and grow to smaller sizes than control group beetles. There were clear and strong effects of larval diet on adult phenotype: an ad libitum pollen diet resulted in paler adult coloration, shorter development time, and larger adult size relative to a limited-availability aphid diet. Our results suggest that the best environment for producing bright-red coloration may not be the best environment for favorable expression of life history characters, especially under stressful conditions. Interactions between different life history stages of H. axyridis are also discussed. Received: 20 April 1997 / Accepted: 30 September 1997     

Effect of different preparatory states on the reflex responses of ankle flexor and extensor muscles to a sudden drop of support surface during standing in humans

The present study hypothesized that if subliminal facilitation of the ankle flexor motoneurons is related to preparatory state to a mechanical perturbation while standing, different standing conditions such as standing with eyes-closed or touching a bar should have different influences on the facilitation levels. While the subjects were maintaining an upright stance posture, sudden drops of support surface were applied several times in the following four different conditions: (1) standing with eyes-open, (2) with eyes-closed, (3) with eyes-open and warning of the drop, and (4) with eyes-open and touching a bar with both hands. The results demonstrated that the EMG responses in the ankle flexor TA muscle increased in the eyes-closed condition, while it reduced in the conditions, such as that the subjects had prior warning or touched a bar. The preparatory state of supraspinal neural centers was suggested to be related to the observed variation in the EMG responses.     

Cuticular receptor activation of postural motoneurons in the abdomen of the hermit crab,<Emphasis Type="Italic"> Pagurus pollicarus</Emphasis>

Displacement of the abdominal cuticle of the hermit crab, Pagurus pollicarus, activates motoneurons of the ventral superficial muscles that mediate posture and slow movements. Five excitatory motoneurons innervating the right ventral superficial muscle of the fourth abdominal segment were activated in a phasic stereotyped fashion in the isolated nervous system. Intracellular records from these motoneurons showed an initial monosynaptic burst, a period of inhibition in which inhibitory post-synaptic potentials were present and then a later period of increased spike frequency generated by excitatory post-synaptic potentials. The reflex response was maintained after severing all ganglionic roots from peripheral structures, isolating the nerve cord from peripheral feedback pathways. The two excitatory components of the response showed a dependence on strain that was much smaller than that found in sensory afferents. There was no relationship between the site of touch to the cuticle and the intensity or pattern of activation of the motoneurons. The reflex burst produced a transient activation of both longitudinal and transverse/circular layers of the muscle with forces that varied between 10% and 25% of the maximum muscle force. These results are consistent with a feedforward regulation of muscle stiffness.     

Relationship between neuromuscular fatigue and spasticity in chronic stroke patients: A pilot study

IntroductionThe aim of this study was to assess the effects of neuromuscular fatigue on stretch reflex-related torque and electromyographic activity of spastic knee extensor muscles in hemiplegic patients. The second aim was to characterize the time course of quadriceps muscle fatigue during repetitive concentric contractions.MethodsEighteen patients performed passive, isometric and concentric isokinetic evaluations before and after a fatigue protocol using an isokinetic dynamometer. Voluntary strength and spasticity were evaluated following the simultaneous recording of torque and electromyographic activity of rectus femoris (RF), vastus lateralis (VL) and biceps femoris (BF).ResultsIsometric knee extension torque and the root mean square (RMS) value of VL decreased in the fatigued state. During the fatigue protocol, the normalized peak torque decreased whereas the RMS of RF and BF increased between the first five and last five contractions. There was a linear decrease in the neuromuscular efficiency-repetitions relationships for RF and VL. The peak resistive torque and the normalized RMS of RF and VL during passive stretching movements were not modified by the fatigue protocol for any stretch velocity.DiscussionThis study showed that localized quadriceps muscle fatigue caused a decrease in voluntary strength which did not modify spasticity intensity. Changes in the distribution of muscle fiber type, with a greater number of slow fibers on the paretic side, may explain why the stretch reflex was not affected by fatigue.