Standing sedation management of a domesticated reindeer for third eyelid removal.

Background: Reindeer are becoming popular animals within petting farms. Few case reports describe the sedation of domesticated reindeer, but none describe the use of ocular local anesthetic blocks in this species. Case Description: A 9-year-old, female, Svalbard reindeer (Rangifer tarandus platyrhynchus) presenting for removal of a squamous cell carcinoma involving the third eyelid. Standing sedation was performed using initial boluses of medetomidine and butorphanol via intramuscular injection before catheter placement and maintenance with a variable rate infusion of medetomidine. Supraorbital, auriculopalpebral, infratrochlear blocks and local infiltration of the base of the third eyelid were performed using mepivacaine. Following the surgical removal of the third eyelid, atipamazole was administered intramuscularly to antagonize the effects of medetomidine. The patient recovered without complications. Conclusion: Medetomidine-butorphanol in combination with local anesthetic blocks provided a sufficient plane of sedation and analgesia for extra ocular surgery in a domesticated reindeer.

Nictitating membrane resection in the horse: a comparison of long-term outcomes using local vs. general anaesthesia.

REASONS FOR PERFORMING STUDY: Neoplasia, for which surgical excision is a frequent treatment, is the most common disease of the equine nictitating membrane. There is little long-term follow-up information available to the practitioner regarding the long-term effects of nictitating membrane excision on ocular health. No information is available to compare recurrence of primary neoplasia of the nictitating membrane after excision with local or general anaesthesia. OBJECTIVES: To evaluate the long-term complications of nictitating membrane resection in horses; recurrence of neoplasia of the nictitating membrane when nictitating membrane resection is performed under local vs. general anaesthesia and if the method of anaesthesia used to permit resection of the affected membrane influences the recurrence of neoplasia of the nictitating membrane after complete nictitating membrane resection. METHODS: Records of 26 horses receiving resection of the nictitating membrane for primary neoplasia of the nictitating membrane 1999-2009 were reviewed. Clinical examination findings, surgical procedure, anaesthesia type, histopathological findings and details of adjunctive treatment were recorded. Owners were contacted via telephone regarding post operative outcomes. Data were analysed using a Fisher's exact test (P<0.05). RESULTS: The most common long-term complication of nictitating membrane excision was mild ocular discharge. Squamous cell carcinoma was the most frequent histopathological diagnosis. Recurrence of neoplasia was uncommon (2/26 horses). No significant difference in the number of horses experiencing recurrence of neoplasia was detected between groups receiving general anaesthesia vs. those receiving local anaesthesia. CONCLUSIONS: Resection of the nictitating membrane in horses following local anaesthesia is not associated with increased risk of recurrence of neoplasia compared with excision under general anaesthesia. Resection of the nictitating membrane is not associated with any long-term ocular side effects and can be an effective modality for cure of primary neoplasia of the nictitating membrane in selected cases.

Treatment of severe keratoconjunctivitis sicca by parotid duct transposition after tympanic neurectomy in rabbits.

PURPOSE: To investigate the feasibility of parotid duct transposition after tympanic neurectomy to treat severe keratoconjunctivitis sicca (KCS) in rabbits. METHODS: Thirty rabbits were divided into three groups in experiment 1. One eye was operated on, and the contralateral eye served as the control. In the KCS group, the lacrimal gland, harderian gland, and nictitating membrane were removed. In the group with parotid duct transposition (DT), the parotid duct was transposed into the lower conjunctival fornix. In the group with parotid duct transposition after tympanic neurectomy (DTTN), the tympanic nerve was resected in addition to parotid duct transposition. Schirmer test was performed and density of corneal staining was determined monthly after surgery, and goblet cell density was measured at postoperative month 3. In experiment 2, the tympanic nerve was resected on one side in 12 rabbits. Both sides of the parotid gland were resected for histopathology at intervals of 2 months to 1 year after surgery. RESULTS: Tear secretion from operated eyes at rest increased significantly after surgery in the treatment groups compared with the KCS group. Tear secretion from operated eyes after chewing was significantly lower in the DTTN than in the DT group. The corneal staining scores were higher in the operated than in the control eyes of the three groups, without significant difference among the operated eyes. Parotid gland atrophy on the operated side occurred at postoperative month 4 and recovered to normal 1 year after surgery. CONCLUSIONS: Parotid duct transposition after tympanic neurectomy could effectively reduce gustatory epiphora but may be insufficient to promote ocular surface health.

Unpaired extinction: implications for treating post-traumatic stress disorder.

Extinction of fear is important for treating stress-related conditions particularly post-traumatic stress disorder (PTSD). Although traditional extinction presents the feared stimulus by itself, there is evidence from both clinical and basic research that repeatedly presenting the feared stimulus by itself does not prevent fear from returning. This renewal or relapse can be "thwarted" by unpaired extinction-presentations of the feared stimulus and the event producing the fear. However, no matter how effective standard unpaired extinction may be in the laboratory, repeated presentation of a traumatic event is untenable. To make an unpaired extinction procedure more clinically relevant, we classically conditioned the rabbit nictitating membrane response using electrical stimulation or air puff as the unconditioned stimulus and then during unpaired extinction reduced both the intensity of the unconditioned stimulus and the days of unpaired stimulus presentations. We found unpaired extinction reduced conditioned and exaggerated unconditioned responding (an animal analog of PTSD called conditioning-specific reflex modification) and could be accomplished with a weak unconditioned stimulus as long as extended presentations were used. Surprisingly, brief presentations of a weak unconditioned stimulus or extended presentations of a strong one made the exaggerated responses stronger. One implication is that brief treatment may not just be ineffectual; it may heighten the symptoms of PTSD. Another implication is that using strong stimuli may also heighten those symptoms.

Scalar timing varies with response magnitude in classical conditioning of the nictitating membrane response of the rabbit (Oryctolagus cuniculus).

The present experiment was aimed at characterizing the timing of conditioned nictitating membrane (NM) movements as function of the interstimulus interval (ISI) in delay conditioning for rabbits (Oryctolagus cuniculus). Onset latency and peak latency were approximately, but not strictly, scalar for all but the smallest movements (<.10 mm). That is, both the mean and standard deviation of the timing measures increased in proportion to the ISI, but their coefficients of variation (standard deviation/mean) tended to be larger for shorter ISIs. For all ISIs, the absolute timing of the NM movements covaried with magnitude. The smaller movements (approximately, .11-.50 mm) were highly variable, and their peaks tended to occur well after the time of US delivery. The larger movements (>.50 mm) were less variable, and their peaks were better aligned with the time of US delivery. These results are discussed with respect to their implications for current models of timing in eyeblink conditioning.

Evidence from retractor bulbi EMG for linearized motor control of conditioned nictitating membrane responses.

Classical conditioning of nictitating membrane (NM) responses in rabbits is a robust model learning system, and experimental evidence indicates that conditioned responses (CRs) are controlled by the cerebellum. It is unknown whether cerebellar control signals deal directly with the complex nonlinearities of the plant (blink-related muscles and peripheral tissues) or whether the plant is linearized to ensure a simple relation between cerebellar neuronal firing and CR profile. To study this question, the retractor bulbi muscle EMG was recorded with implanted electrodes during NM conditioning. Pooled activity in accessory abducens motoneurons was estimated from spike trains extracted from the EMG traces, and its temporal profile was found to have an approximately Gaussian shape with peak amplitude linearly related to CR amplitude. The relation between motoneuron activity and CR profiles was accurately fitted by a first-order linear filter, with each spike input producing an exponentially decaying impulse response with time constant of order 0.1 s. Application of this first-order plant model to CR data from other laboratories suggested that, in these cases also, motoneuron activity had a Gaussian profile, with time-of-peak close to unconditioned stimulus (US) onset and SD proportional to the interval between conditioned stimulus and US onsets. These results suggest that for conditioned NM responses the cerebellum is presented with a simplified "virtual" plant that is a linearized version of the underlying nonlinear biological system. Analysis of a detailed plant model suggests that one method for linearising the plant would be appropriate recruitment of motor units.

Selective attention and Pavlovian conditioning.

The present results show that the common practice of using self-indexing conditioned stimuli (CSs) in research on Pavlovian conditioning is a major source of experimental bias. The typical stimulus used is either a light flash or a sound pulse in a light/sound-shielded chamber. Under these conditions the onset characteristics of the CS signal totally predominate over the durational characteristic, i.e. the pattern information. Thus a visual pattern presented as a CS in a dark chamber is confounded with a brightness change from darkness to light. In the first experiment, animals were conditioned with a brightness CS using a self-indexing signal paradigm. When tested for specificity of the conditioning, they showed complete transfer of learning to either a visual pattern or even an auditory CS. These findings indicated that the traditional conditioning paradigm is biased towards non-specific sensory learning. The second experiment showed that specific sensory conditioning is critically dependent on selective attention mechanisms. When the onset characteristics of the CS signal were de-emphasized by the use of equal energy background illumination in the intertribal interval (ITI) during conditioning, the animals were not able to feature extract either the onset or the durational component of the CS signal from the ITI background despite prolonged training. It was only by starting with conditioning that was initially anchored to the CS onset characteristics that a perceptual fade-in procedure would bias attention to feature extract the durational characteristics of the CS. Thus conditioning occurred only when the rabbit's attention was directed to detection of the gratings display without any associated changes in visual albedo. Perhaps the most important finding of the present experiments is that the use of self-indexing CS signals in Pavlovian conditioning inevitably introduces non-specific sensory processing involving multiple sensory input pathways in the conditioning. This inherent uncertainty of the sensory input pathways presents a problem for clarifying the role of sensory pathways in the neural mechanisms of NM conditioning. In addition, the use of self-indexing CSs inevitably leads to an underestimation of the role of forebrain mechanisms in Pavlovian conditioning.

The effect of rewarding hypothalamic stimulation on behavioral and neural hippocampal responses during trace eyeblink conditioning in rabbit (Oryctolagus cuniculus).

Rabbits were trace-conditioned with a tone as a conditioned stimulus and an airpuff as an unconditioned stimulus. Electrical stimulation to the medial forebrain bundle in the lateral hypothalamus was delivered either before or after the tone-airpuff pair. The purpose of the present study was to test whether the effect of post-trial hypothalamic stimulation differed from the effect of pre-trial hypothalamic stimulation on trace conditioning in the same subjects. Additionally, hippocampal responses were measured during sessions to see if hypothalamic stimulation activated dopaminergic fibres and affected hippocampal cell functioning and thus learning. The results showed that behavioral nictitating membrane conditioned responses were acquired quickly and hippocampal multiple unit activity increased with post-trial hypothalamic stimulation in comparison to almost non-existent conditioned responses and activity changes in the pre-trial hypothalamic stimulation sessions. The results suggest that hypothalamic stimulation affects trace conditioning differently depending on its time of delivery during conditioning.

Learned modification of the nictitating membrane reflex by auditory stimuli in the barn owl.

The barn owl, Tyto alba, extends its nictitating membrane (NM) in response to an air puff to the cornea or a mild para-orbital electrodermal shock. The NM extension habituated rapidly if the stimulus was repeated. Habituation was prevented by pairing the aversive stimulus with a sound. The sound stimulus did not, by itself, induce an NM extension. Repeated pairing of sound with the aversive stimulus caused the subjects to modify the duration of their NM extension, increasing the duration when exposed to longer aversive stimuli and decreasing in response to shorter stimuli. No transference of the response was seen from the aversive stimulus to the sound. The learned change in duration of the NM extension resisted extinction. This modification of the NM extension reflex resembles previous descriptions of primer-produced facilitation.

Neuronal activity in the medial prefrontal cortex during Pavlovian eyeblink and nictitating membrane conditioning.

The present study assessed Pavlovian eyeblink (EB) conditioning, using tones and periorbital shock as the conditioned and unconditioned stimuli (CS and US), and nictitating membrane (NM) conditioning, using tones and airpuffs as the CS and US. During each experiment, CS-evoked changes in multiple-unit activity (MUA) in the medial prefrontal cortex (mPFC) were recorded. Concomitant heart rate (HR) conditioned responses (CRs) were also recorded. A nonassociative control group received explicitly unpaired presentations of the CS and US in each experiment. Increases in both NM and EB CRs occurred over sessions in the paired, but not the unpaired, groups. Decelerative HR CRs also occurred in the eyeshock, but not the airpuff, group. Although tone-evoked increases in neuronal activity were obtained during 10 initial tone-alone presentations in all groups, this activity habituated over trials. CS-evoked increases in neuronal activity also occurred, but this activity was considerably greater in the group that received periorbital shock as the US. During subsequent extinction trials, decreases in tone-evoked neuronal activity occurred in this group, compared with the previous CS/US paired trials. CS-evoked MUA increases were minimal during all except the pretraining phase of the study in the CS/US unpaired control groups and in the paired airpuff group. These findings show that neuronal activity during associative learning occurs in the mPFC during Pavlovian EB, as well as HR conditioning, but this activity apparently reflects an affective component to learning that is only indirectly related to skeletal conditioning.

Nimodipine facilitates retention of the classically conditioned nictitating membrane response in aged rabbits over long retention intervals.

Aged rabbits initially underwent 18 days of acquisition of the classically conditioned nictitating membrane response (NMR) using a tone conditioned stimulus (CS) and an air puff unconditioned stimulus (UCS). They were then treated with a low or high dose of nimodipine or a vehicle for 90 days. During this time no further CS-UCS pairings were presented. They underwent testing for retention of the conditioned response (CR) at 30 and 90 days. Retention testing consisted of 20 presentations of the CS alone. Rabbits in the control condition retained 46.4% of their predrug levels of conditioned responding and rabbits receiving the low dose of nimodipine retained 37.3% of their predrug levels after 30 days. After 90 days, retention in these animals declined to 8.1% and 14.1%, respectively. In contrast, rabbits receiving the high dose of nimodipine retained 85% of their predrug learning at 30 days with little decline at 90 days (77.1%). Nonassociative factors such as sensitivity to the CS or UCS could not explain these effects.

Treatment of canine nictitans plasmacytic conjunctivitis with 0.2 per cent cyclosporin ointment.

Nictitans plasmacytic conjunctivitis, commonly referred to as plasma cell infiltrate of the nictitans or plasmoma, was diagnosed in 12 dogs (23 eyes) on the basis of clinical signs and nictitans conjunctival biopsy specimens. These dogs underwent a clinical therapeutic trial with twice daily 0.2 per cent cyclosporin ophthalmic ointment. Response to therapy was monitored over a six-week period and repeat biopsy specimens were then taken. Significant (P < 0.05) reductions between pre- and post trial scores were recorded for: mucopurulent ocular discharge quantity; degree of bulbar conjunctival hyperaemia; areas of nictitans hyperaemia, thickening and depigmentation. Schirmer tear test values significantly increased between the start and end of treatment. Biopsy specimens were subjected to selective detection procedures for plasma cells (methyl green-pyronin staining) and T lymphocytes (CD3 antigen labelling). Mean cell counts showed a significant reduction in plasma cell numbers, but the trend towards reduced T lymphocyte numbers was not significant.

Quantitative distribution of protein kinase C alpha, beta, gamma, and epsilon mRNAs in the hippocampus of control and nictitating membrane conditioned rabbits.

We used oligonucleotide in situ hybridization and film autoradiography to quantitate the distributions of protein kinase C (PKC) alpha, beta, gamma, and epsilon mRNAs in subregions of rabbit hippocampus. Levels of each of the hippocampal PKC isozyme mRNAs and patterns of their regional distributions were remarkably invariant between individuals. Within stratum pyramidale, the highest levels of PKC alpha mRNA were in the CA2 region, while PKC beta mRNA was maximally expressed in CA1, and PKC epsilon mRNA in CA3; PKC gamma mRNA was abundantly expressed throughout Ammon's horn. Previous experiments employing quantitative autoradiography for [3H]PDBU (Olds et al., Science, 245 (1989) 866-869) revealed an increase in membrane-bound PKC in the CA1 region of rabbit hippocampus up to 3 days following classical conditioning of the nictitating membrane response. We report here that there were no differences in levels of PKC alpha, beta, gamma, or epsilon mRNA between conditioned and control rabbits in any hippocampal region one day after training. These data are consistent with the hypothesis that PKC is post-translationally activated and translocated to the membrane during memory storage.

Muscle activity during unconditioned and conditioned eye blinks in the rabbit.

EMGs were recorded from the orbicularis oculi, retractor bulbi and superior rectus muscles in rabbits to investigate the time course of muscle activation during unconditioned and conditioned eye blinks. EMGs from the three muscles showed two responses, with the responses of the orbicularis oculi and retractor bulbi showing the same latency, and the responses of the superior rectus lagging. The latency of responses to periorbital electrostimulation was about 5 ms, and to air puff stimulation about 10 ms. Results showed a tight coupling of activity between muscles, with cross-correlograms peaking at 0.65 to 0.85 and showing little time shift. Stimulus-response curves showed clear non-linearities in the response of the muscle to changes in stimulus strength. Local anesthesia of the cornea had little effect on unconditionally evoked responses. The form of unconditionally evoked responses was similar with periorbital electrostimulation and air puff stimuli but differed in latency. These results show the form of the eye blink reflex response and will be of importance in interpreting electrophysiological studies of the classically conditioned eye blink of rabbits.

Cerebellar cortex and eyeblink conditioning: a reexamination.

We examined the effects of cerebellar cortical lesions upon conditioned nictitating membrane responses in rabbits. Using extended postoperative conditioning and unpaired presentations of the conditioned stimuli (CSs), we confirmed that combined lesions of lobules HVI and ansiform lobe abolished conditioned responses (CRs) established to light and white noise CSs. Extended retraining enabled some slight recovery of CR frequencies. Less extensive cortical lesions produced initial abolition of CRs but allowed more complete recoveries. Although CR frequencies and amplitudes were profoundly depressed by cortical lesions, unconditioned response (UR) amplitudes to periorbital electrical stimulation were enhanced. The dissociation of lesion effects upon conditioned and unconditioned responses is consistent with the suggestion that cerebellar cortical mechanisms are important for the learning and execution of eyeblink conditioning.

Comparison of peripheral and central nervous system sympatholytic actions of prazosin using the cat nictitating membrane.

Prazosin is a highly selective alpha 1-adrenoceptor antagonist that decreases blood pressure by actions on both the peripheral and central (CNS) divisions of the nervous system. The present investigation was undertaken in an attempt to characterize the relative contribution of these two sympatholytic sites of action. Submaximal contractions of the nictitating membranes were evoked by electrical stimulation of the preganglionic cervical sympathetic nerve trunk and by stimulation of the posterior hypothalamus in anesthetized cats. In initial control experiments, phenoxybenzamine (0.1-3.0 mg/kg i.v.) produced an equivalent depression of evoked nictitating membrane responses from both peripheral and CNS sites of activation which suggests only a peripheral blocking action as well as functional equivalence of the intensity of CNS and peripheral nerve stimulation. In contrast, prazosin (3-300 micrograms/kg i.v.) caused a differential dose-related depression of the evoked responses with ED50s of 81.5 micrograms/kg (peripheral stimulation) and 12.5 micrograms/kg (CNS stimulation) respectively; P less than 0.05. Pretreatment with rauwolscine (500 micrograms/kg i.v.) totally prevented the differential CNS sympatho-inhibition produced by prazosin. These results indicate that, although both CNS and peripheral sites of drug action are manifest, the ED50 for prazosin-induced CNS sympatho-inhibition is approximately 6-fold less than that required for direct alpha 1-adrenoceptor blockade at the end organ. In addition, prazosin produces CNS sympatho-inhibition indirectly by means of an alpha 2-adrenoceptor mechanism.

Conditioned inhibition of the nictitating membrane response in rabbits following hypothalamic and mesencephalic lesions.

Rabbits were trained on a Pavlovian conditioned inhibition (CI) task using light as the reinforced conditioned stimulus (CS+) and the same light compounded with a tone as the nonreinforced CS-. The conditioned response was the nictitating membrane response. After attaining a criterion of CI performance, animals received radio-frequency lesions of the hypothalamus (n = 11) or midbrain (n = 14). For the hypothalamic lesion cases, primary damage extended from the optic chiasm to the pretectal region. For the mesencephalic lesion cases, primary damage ranged from the most rostral portions of the periaqueductal grey (PAG) caudally to the tegmental reticular formation at the level of the third nerve. Prior research suggested that the hypothalamic lesions would disrupt retention of CI by increasing responding to the CS-. Except where a lesion impinged upon the zona incerta, no CI disruption was observed. In accordance with previous studies (Berthier, N.E. and Moore, J.W., Physiol. Behav., 25 (1980) 667-673; Mis, F.W., J. Comp. Physiol. Psychol., 91 (1977) 975-988), post-lesioning CI disruption was observed in some of the mesencephalic lesion cases involving the posterior commissure, PAG and/or accessory oculomotor nuclei. However, CI performance recovered over the course of retraining.

Single-unit activity in red nucleus during the classically conditioned rabbit nictitating membrane response.

Previous investigations have suggested that the cerebellum and associated brainstem structures, including the red nucleus, are essential for the expression of the classically conditioned nictitating membrane (NM) response. The present study examined the firing patterns of extracellularly-recorded single units in the red nucleus of the awake rabbit during differential conditioning. Tones were used as conditioned stimulus (CS+ and CS-) and periocular electrostimulation was used as the unconditioned stimulus (US). Most units exhibited one or more changes in firing rate during the presentation of the CS, and increases in firing were much more common than decreases. The onset of some of these changes appeared to be time-locked to the onset of the CS ('CS-locked' responses), while other changes were time-locked to the onset of the CR ('CR-locked' responses). About one-third of all CS-locked changes were CR-dependent, meaning that the neuronal response was reduced when the CR did not occur. About two-thirds of all CR-locked responses preceded the onset of the CR, and lead times varied considerably across units. Many CR-locked units were located in what has been described as a dorsal face region of the red nucleus. Most units responded to the US, and some of the US responses were CR-dependent: i.e., a smaller US response was evoked when a CR preceded the US than when the CR was absent. Our results support the notion that cerebellum-brainstem circuits are involved in generating NM CRs.

Reflex facilitation of the nictitating membrane response remains after cerebellar lesions.

Reflex facilitation and associated properties were investigated during classical conditioning of the nictitating membrane (NM) response in rabbit. In the first experiment, the role of the cerebellum was examined by comparing the unconditioned responses of animals with bilateral lesions of the deep cerebellar nuclei with those of operated controls during counterbalanced tone/light (T/L) discrimination training. Both T and L facilitated unconditioned NM responses when used as the CS+ (conditioned stimulus), but neither facilitated when used as the CS-. There were no significant differences in the amount of reflex facilitation exhibited by animals with lesions compared with control animals. Animals with lesions, however, failed to acquire conditioned responses after 10 days of training, whereas all control animals met acquisition criterion within 4 days. In the second experiment, reflex facilitation was shown to decrement in a stimulus-specific manner when nonreinforced presentations of an auditory stimulus were given. The discussion of results focuses on the relation between reflex facilitation and classical conditioning in terms of behavioral properties and underlying neural systems.